阅读说明：本技术 用于治疗移植物抗宿主疾病的组合物 (Compositions for the treatment of graft versus host disease ) 是由 让·德冈兹伯格 于 2019-08-05 设计创作，主要内容包括：本发明涉及用于治疗或预防移植物抗宿主疾病的组合物和方法。(The present invention relates to compositions and methods for treating or preventing graft versus host disease.)

1. A substance selected from the group consisting of an adsorbent and an antibiotic inactivating enzyme for use in a method of treating, preventing, delaying or reducing the risk or severity of graft-versus-host disease in a subject.

2. The substance for use according to claim 1, wherein the subject to be treated receives, will receive or has received an agent that causes dysbiosis.

3. The substance for use according to claim 2, wherein the agent causing dysbiosis is an antibiotic administered for the treatment or prevention of infection.

4. The substance for use according to any one of claims 1 to 3, wherein the substance is an adsorbent.

5. The substance for use according to claim 4, wherein the adsorbent is activated carbon.

6. The substance for use according to claim 4 or 5, wherein the adsorbent is in a formulation comprising:

-a core containing said adsorbent, and

-an outer coating layer formed around the core such that the adsorbent is released from the formulation in the lower part of the intestine.

7. The substance for use according to claim 6, wherein the core further comprises a carrageenan, such as kappa-carrageenan.

8. Substance for use according to any one of claims 1 to 3, wherein the substance is an antibiotic inactivating enzyme, wherein the subject receives, will receive or has received an antibiotic for the prevention or treatment of an infection, and wherein the enzyme is adapted to inactivate the antibiotic.

9. The substance for use according to claim 8, wherein (i) the enzyme is a beta-lactamase, in particular VIM-2 or ribaxamase, and the antibiotic is a beta-lactam antibiotic; or (ii) the enzyme is an erythromycin esterase and the antibiotic is a macrolide.

10. The substance for use according to claim 8, wherein the enzyme is a hybrid protein molecule comprising two antibiotic-inactivating enzymes bonded together, the two enzymes inactivating the same or different antibiotics or antibiotics from the same or different classes.

11. Substance for use according to any one of claims 8 to 10, formulated in a composition for oral administration suitable for releasing the antibiotic inactivating enzyme in a desired part of the intestine, such as the lower part of the intestine.

12. Substance for use according to any one of claims 2 to 11, wherein the agent causing dysbiosis is an antibiotic selected from the group consisting of:

- β -lactams comprising:

penicillins (e.g., penicillin G, penicillin V, ampicillin, amoxicillin, bacampicillin, carbenicillin, ticarcillin, azlocillin, mezlocillin, piperacillin, etc.),

penicillinase-resistant penicillins (e.g., methicillin, oxacillin, cloxacillin, dicloxacillin, nafcillin, etc.),

cephalosporins, for example: first generation cephalosporins (e.g. cefadroxil, cephalexin, cephradine, cephalothin, cefapirin, cefazolin, etc.); second generation cephalosporins (e.g., cefaclor, cefamandole, cefonicid, cefoxitin, cefotetan, cefuroxime axetil, cefmetazole, cefprozil, chlorocefcapene, cefradide, etc.); third-generation cephalosporins (e.g., cefepime, cefoperazone, cefotaxime, ceftizoxime, ceftriaxone, ceftazidime, cefixime, cefpodoxime, ceftibuten, etc.); fourth generation cephalosporins (e.g. cefixdine, cefepime, cefozopran, cefpirome, cefquinome, etc.); fifth and other generation cephalosporins (e.g. cefepime, ceftaroline, ceftizolidine, etc.),

carbapenems (e.g. imipenem, meropenem, ertapenem, doripenem, etc.),

monocyclic lactams (e.g. aztreonam and the like),

quinolones (e.g. nalidixic acid) and fluoroquinolones (e.g. cinoxacin, ciprofloxacin, moxifloxacin, levofloxacin, ofloxacin, gatifloxacin, gemifloxacin, norfloxacin etc.),

sulfonamides (e.g. sulfanilamide, sulfadiazine, sulfamethoxazole, sulfisoxazole, sulfacetamide, sulfamethoxydiazine, etc.),

aminoglycosides (e.g. streptomycin, gentamicin, tobramycin, amikacin, netilmicin, kanamycin, neomycin B, C and E, spectinomycin, puromycin, gentamicin and the like),

tetracyclines (e.g. tetracycline, chlortetracycline, oxytetracycline, methacycline, doxycycline, minocycline, tigecycline, elacycline, etc.),

macrolides (e.g. erythromycin, azithromycin, clarithromycin, fidaxomycin, telithromycin, josamycin, oleandomycin, spiramycin, tylosin, roxithromycin, quinomycin, solithromycin, etc.),

glycopeptides (e.g. vancomycin, oritavancin, telavancin, teicoplanin, dalbavancin, ramoplanin, etc.),

oxazolidinones (e.g. linezolid, epsiprazole, tedizolid, ralazolide, cycloserine, etc.),

chloramphenicol (e.g., chloramphenicol, thiamphenicol, etc.),

lincosamides (e.g. clindamycin, lincomycin, etc.),

streptogramins (e.g. pristinamycin, quinupristin/dalfopristin, virginiamycin, etc.),

polymyxins (e.g. polymyxin A, B, C, D, E1 (colistin A) or E2, colistin B or C etc.),

diaminopyrimidines (e.g., trimethoprim, pyrazinamide, etc., commonly used in combination with sulfamethoxazole),

sulfones (e.g. dapsone, sodium aclidinesulfonate, etc.),

-a source of para-aminobenzoic acid,

-a bacitracin which is capable of inhibiting the growth of bacillus,

-a source of isoniazid,

rifampins (e.g. rifampin, rifabutin, rifapentine, rifalazil, rimamixin, etc.),

-a source of-ethyl-amine-butanol,

-a source of N-methyl-aminothioisonicotinamine,

-capreomycin, and

-clofazimine.

13. Active compound for use according to any one of claims 1 to 12, wherein the subject is or may be a recipient of allogeneic cells, tissues or organs.

14. An active compound for use according to any one of claims 1 to 13, wherein the subject is or may be a recipient of hematopoietic stem cells.

Technical Field

The present invention relates to compositions and methods for treating and/or preventing Graft Versus Host Disease (GVHD). In particular, the invention is useful in preventing antibiotic-induced destruction of the gut microflora and reducing or preventing the development of GVHD, particularly in humans, in patients who receive or will receive potentially immunocompetent transplants, such as allogeneic hematopoietic stem cell transplantation.

Background

Graft Versus Host Disease (GVHD) is a major cause of graft-related morbidity and mortality. GVHD can occur after immunocompetent transplantation. In particular, GVHD may occur following allogeneic cell transplantation, such as stem cell transplantation and/or bone marrow transplantation. GVHD can also occur after transfusion. In GVHD, the transplanted immune cells can recognize the host's cells as foreign and attack them. Patients with GVHD may often have symptomatic manifestations of three organs such as skin (rash/dermatitis), liver (hepatitis/jaundice), and gastrointestinal tract (abdominal pain/diarrhea). Symptoms can be very significant and may even cause death of the host.

Graft-versus-host disease can be largely classified into acute graft-versus-host disease and chronic graft-versus-host disease. GVHD is acute when it begins within 100 days after allogeneic hematopoietic stem cell transplantation, and is usually the response of the donor immune cells to host tissue. Activated donor T cells typically damage host epithelial cells following the inflammatory cascade following initiation of the priming protocol. Statistically, about 35% to 50% of recipients/patients of Hematopoietic Stem Cell Transplantation (HSCT) are likely to develop acute GVHD. The specific risk generally depends on the stem cell source, the age of the patient, the formulation and the GVHD prevention/treatment used. Acute GVHD is usually staged and graded (grade 0-IV) according to the number and extent of organs involved. Patients with grade III/IV acute GVHD tend to have adverse outcomes with high mortality. Chronic GVHD is the most major and common side effect after HSCT, occurring in 20% -70% of patients who live 100 days after blood and bone marrow progenitor cell transplantation, and is the leading cause of post-transplant death.

Currently, standard GVHD prevention regimens are non-specific: immunosuppression is performed by targeting the proliferative process of immune cells, such as methotrexate, tacrolimus, everolimus, sirolimus, mycophenolate mofetil, or cyclosporine a. In addition, GVHD treatment typically involves the administration of corticosteroids. However, since these immunosuppressive agents have non-specific effects, their toxicity is very high, and the damaged immune system becomes very sensitive to infectious diseases such as bacterial infections.

Prior to any HSCT, recipients were placed under a preparation regimen (conditioning phase) which placed them in a neutropenic state, thus making them very susceptible to a variety of different infections. In the case of fever, when there is a fear of bacterial infection, the patient will be treated with antibiotics to calm the infection at the earliest stage. Regardless of whether the antibiotic is administered orally or parenterally, a significant portion of the administered agent reaches the colon in an active form, where it comes into contact with the gut microflora, a large number of commensal bacteria living in the gastrointestinal tract, primarily the caecum and the colon. As a result of this process, the composition of the microbial flora is profoundly altered, affecting both anaerobic bacteria (which perform a major physiological role in the intestines of normal subjects and animals) and aerobic bacteria.

Dysbiosis (dysbiosis) or an imbalance in the gut microflora has been associated with many diseases, including GVHD, in HSCT recipients. The relationship between microbial flora and GVHD has long been suspected but is still not fully understood. Some recent work demonstrated a direct correlation between loss of microbiome diversity and an increase in GVHD severity (Holler et al, 2014.Biology of Blood and Marrow transfer 20(5): 640-. Furthermore, more direct evidence for the role of the gut microflora in the development of GVHD comes from studies conducted in patients that show a correlation between an increased abundance of commensal bacteria, particularly of the genus Blautia (Blautia), and a reduction in the incidence of fatal GVHD in those patients (Jenq et al, 2015.Biology of Blood and Marrow transfer 21(8): 1373-1383).

It has also been recently shown that the use of antibiotics, particularly broad spectrum antibiotics, leads to increased severity of GVHD and intestinal injury (Shono et al, 2016.Science relative Medicine 8(339):339ra 71). Furthermore, recent studies have highlighted the deleterious effects of antibiotics in the severity and overall survival of acute GVHD (Routey et al, 2017 OncoImmunology 6(1): e 1258506).

However, in order to prevent and cure bacterial infections in transplanted subjects, the use of antibiotics is necessary. Therefore, there is a high unmet medical need for a pharmaceutically effective solution that allows the use of antibiotics and the prevention and/or treatment of GVHD in this patient population.

Disclosure of Invention

The present invention is based on the observation that GVHD is associated with major changes in the gut microflora that occur following administration of immunocompetent grafts, such as HSCT and antibiotic treatment, suggesting that antibiotic-associated dysbiosis may be one of the causes of the initial appearance and/or severity of GVHD. Antibiotics are not the only agents that may cause dysbiosis. Thus, it is also desirable to prevent the secondary effects of such agents.

Thus, the present invention provides, among other things, compositions and methods for treating or preventing GVHD or reducing the risk thereof in a subject in need thereof based on the use of a substance suitable for inactivating an agent causing dysbiosis.

In certain embodiments, the substance is an adsorbent. In other embodiments, wherein the agent that causes dysbiosis is an antibiotic, the substance is an antibiotic-inactivating enzyme.

Thus, in one embodiment, the invention relates to an adsorbent for use in a method of treating or preventing GVHD or reducing the risk or severity of GVHD in a subject.

The invention also relates to an antibiotic-degrading enzyme for use in a method of treating or preventing GVHD or reducing the risk or severity of GVHD in a subject.

The subject (alternatively referred to as a "host") may be a recipient of allogeneic cells, tissues or organs such as cord blood, bone marrow, peripheral blood, stem cells (e.g., hematopoietic stem cells and adult or embryonic stem cells), blood products, and solid organs potentially containing immunocompetent cells.

In certain embodiments, the subject is a recipient of potential immunocompetent transplants, such as Hematopoietic Stem Cell (HSC), bone marrow, Peripheral Blood (PBSC), and cord blood transplants.

In certain embodiments, the potentially immunologically active transplant is an umbilical cord blood transplant. In certain embodiments, the cord blood transplant is selected from the group consisting of a single cord blood transplant, a double cord blood transplant, a multiple cord blood transplant, a manipulated cord blood transplant, and combinations thereof. In certain embodiments, the manipulated cord blood transplant comprises an ex vivo expanded cord blood transplant. In certain embodiments, the manipulated cord blood transplant comprises treating the cord blood with a prostaglandin prior to transplantation. In certain embodiments, the manipulated cord blood transplant comprises stripping T-cells from the cord blood prior to transplantation.

In certain embodiments, the potentially immunocompetent transplant is a bone marrow transplant. In certain embodiments, the immunocompetent transplant is a peripheral blood transplant. Suitable bone marrow or peripheral blood may be obtained from children or adults. The bone marrow and peripheral blood may be manipulated in any suitable manner prior to transplantation.

In certain embodiments, the potentially immunocompetent transplant is a stem cell transplant. In certain embodiments, the stem cell transplant is an allogeneic stem cell. In certain other embodiments, the stem cell is from an adult.

In certain embodiments, the potentially immunocompetent transplant is an embryonic stem cell transplant. In certain embodiments, the potentially immunocompetent transplant is an organ transplant. In certain embodiments, the potentially immunologically active transplant corresponds to infusion of a blood product. Infusions of blood products may include, but are not limited to, blood, serum, plasma, and platelet infusions and infusions of derivative products.

In particular embodiments, the potentially immunocompetent transplant is not a human embryonic stem cell.

In another specific embodiment, the subject is an immunocompromised subject. In other particular embodiments, the subject is immunocompromised through the effect of immunosuppressive therapy or due to disease, such as immunodeficiency caused by bacterial or viral infection, such as acquired immunodeficiency syndrome (AIDS) which may be the result of infection with Human Immunodeficiency Virus (HIV).

In one aspect, the agent is administered in a therapeutically effective amount such that at least one symptom or characteristic of GVHD is inhibited, or reduced in intensity, severity, duration or frequency, or delayed in onset. In certain embodiments, the at least one symptom or characteristic of GVHD is selected from the group consisting of liver injury, rash, jaundice, intestinal inflammation, mucosal sloughing, diarrhea, abdominal pain, nausea, and vomiting. In certain embodiments, the GVHD is acute GVHD. In certain embodiments, the GVHD is chronic GVHD. In certain embodiments, the subject to be treated is an immunocompromised subject susceptible to receiving a transplant.

In other particular embodiments, the agent is administered in a therapeutically effective amount for a sufficient time to ensure that GVHD, or at least one symptom thereof, is inhibited, or reduced in intensity, severity, duration, or frequency, or delayed in onset. In particular, the substance may be administered periodically, for example once daily, twice daily, three times daily or more than three times daily. For example, the substance may be administered to the subject simultaneously with the dysbiosis-causing agent for the same total time or for several days. Illustrative treatment procedures include, for example, treatment of the agent causing the dysbiosis within 7 days, wherein the substance is also administered during the 7 day exposure to the agent. In another embodiment, the substance may be administered one or more days before and/or after the start of therapy with the dysbiosis causing agent to ensure that most of the residual agent is eliminated. For example, the substance may be administered first on the day before the first day of administration of the agent and for two days after the last day of administration of the agent.

In the context of the present invention, the substance may be an adsorbent, which may be used to adsorb and thus remove from the intestine any residual agent or metabolite thereof which would otherwise cause adverse effects in the host upon reaching the lower intestine and/or colon following oral or parenteral administration of the agent.

In particular embodiments, the subject has received, receives, or will receive an antibiotic for treating the infection. In this embodiment, the substance is administered in order to prevent adverse effects of the antibiotic on the intestinal microflora, in particular in the lower part of the intestine, such as the terminal ileum, the caecum or the colon.

In another specific embodiment, the subject is not receiving antibiotic therapy. In this case, the adsorbent is administered to prevent destruction of the gut flora due to other reasons than antibiotic administration. For example, the adsorbents may be used to treat infections caused by harmful bacteria, such as Clostridium difficile (Clostridium difficile), by affecting the germination or growth of the harmful bacteria, by preventing the production of toxins, or by adsorbing toxins released by these harmful bacteria. In particular, the adverse effects in the host may be caused by any other molecule or toxin that may have serious adverse effects on the gut microflora or intestinal tissue, such as, but not limited to, bacterial toxins and molecules released into the gastrointestinal tract, including those produced by pathogenic microorganisms.

The substance may be formulated for delivery to a desired portion of the intestine. In order to reduce the concentration of antibiotics or other molecules having local adverse effects on the intestinal or intestinal microflora, it may be preferred to release the substance as soon as possible after the absorption of the antibiotic is complete, and a rapid release is preferred. Ideally, the dosage form of the substance is selected to be sufficient to significantly reduce the concentration of the undesirable agent (e.g., antibiotic, other drug or bacterial or fungal toxin) in the intestine and also so that the substance remains effective when released. Representative dosage forms include capsules, tablets, pellets, and other suitable dosage forms that provide a relatively rapid effect on removal of the agent in the colon before the unwanted agent can disrupt the intestinal microbial flora.

Detailed Description

As used herein, the term "acute" when used in conjunction with tissue injury and related diseases, disorders, or conditions has the meaning understood by any person skilled in the medical arts. For example, the term generally refers to the presence of a sudden or severe onset of symptoms in a disease, disorder, or condition. In certain embodiments, the acute injury is caused by an ischemic or traumatic event. Generally, the term "acute" is used in contrast to the term "chronic".

As used herein, the term "chronic" has the meaning as understood by any person skilled in the medical art when used in connection with tissue damage and associated diseases, disorders or conditions. In general, the term "chronic" refers to a disease, disorder or condition that involves persistent and/or recurrent symptoms. Chronic diseases, disorders or conditions typically develop over a long period of time. The term "chronic" is used in contrast to the term "acute". In certain embodiments, the chronic disease, disorder, or condition is caused by cellular degeneration. In certain embodiments, the chronic disease, disorder, or condition is caused by age-related cellular degeneration.

As used herein, the term "preventing," when used in conjunction with the occurrence of a disease, disorder, and/or condition, refers to reducing the risk of the occurrence of the disease, disorder, and/or condition.

In the context of the present invention, a "risk" of a disease, disorder and/or condition includes the likelihood that a particular individual will develop the disease, disorder and/or condition (e.g., GVHD). In certain embodiments, the risk is expressed as a percentage. In certain embodiments, the risk is from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90 up to 100%. In certain embodiments, the risk is expressed as a risk relative to a risk associated with a reference sample or a set of reference samples. In certain embodiments, the reference sample or set of reference samples has a known risk of a disease, disorder, condition, and/or event (e.g., GVHD). In certain embodiments, the reference sample or set of reference samples is from an individual comparable to a particular individual. In certain embodiments, the relative risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or higher.

As used herein, the term "immunocompetent transplant" refers to a tissue transplant obtained from a donor that contains immunocompetent cells of the donor or contains immunocompetent cells of the donor that can differentiate into immune cells. The transplanted immunocompetent cells, such as T cells, may attack and destroy host cells and thereby cause GVHD.

GVHD

GVHD is well known in the art (see, e.g., Ferrara, J. et al, "Graft-versus-host disease" (Graft-cover-host disease), Lancet.2009; 373(9674): 1550-61; MacMillan, M. et al, "analysis of Acute Graft-versus-host disease after unrelated donor umbilical cord Blood transplantation: risk factors" (assay Graft-cover-host disease after unrelated patient umbilical cord Blood transplantation), Blood,2009,113, 11: 2410-2415; Matsumura, T. et al, "Allogeneic umbilical cord Blood transplantation for adult Acute lymphoblastic leukemia: retrospective survey of Japanese patients" (Clinical laboratory of adult umbilical cord Blood, Clinical laboratory of Clinical laboratory of adult umbilical cord Blood, 20117. et al, "Clinical laboratory of adult umbilical cord Blood transplantation for adult umbilical cord Blood, Clinical laboratory of Clinical blod transfer for 30 adlets with pharmaceutical magnetronics), Anticancer Res.,2009(5): 1763-70). The immune system of an individual functions by recognizing certain cell surface proteins, some of which are known as major histocompatibility complex proteins or MHC proteins. There are additional minor histocompatibility proteins that may also contribute to immune recognition events. The immune system of an individual mammal can recognize the MHC proteins of its own or its twin as self and thus not destroy its own cells or its twin cells. Members of the same species may share major and/or minor histocompatibility antigens, and thus an individual may not recognize cells of another member of its species as non-self, depending on the degree of difference between the MHC proteins of the two individuals. When the immune system of an individual recognizes cells of other members of the same species as non-self, the immune system of the first individual may in turn destroy the cells of the second individual. In humans, the major histocompatibility proteins are referred to as "HLA" antigens.

When tissue, such as bone marrow, blood cells, or a solid organ, is transplanted from one individual to another, the recipient may recognize the donor's cells as non-self and the recipient's immune system may destroy the donor's cells. Thus, in tissue transplantation, the recipient may be subjected to immunosuppressive drugs and/or irradiation. However, in GVHD, the transplant patient may also experience immune recognition in the opposite direction, i.e., the donor's tissue may contain immunocompetent cells, which in turn destroy the recipient's cells. In certain cases, when a patient receives an HSCT transplant from a healthy donor, the patient's immune system is first ablated by radiation or chemotherapy, due to the possible presence of harmful, usually malignant cells as a result of the transplant. In this case, the transplanted immune system may recognize the patient's cells as non-self and attack them.

GVHD can occur when any allogeneic cell, such as cord blood, bone marrow, peripheral blood, adult stem cells, embryonic stem cells, blood products, and/or solid organs containing immunocompetent cells, is transferred from a donor to a recipient. Thus, when there is a difference in MHC antigenicity between the donor and recipient, the recipient is at risk of developing GVHD. T-lymphocytes from the donor recognize the difference on the basis of HLA antigens and attack the new body, the recipient's body, although the HLA markers of most patients and donors match as closely as possible. In particular, GVHD is caused when immunocompetent T cells in a donor transplant are infused into an immunocompromised recipient. However, many secondary markers differ from donor to patient except when the patient and donor are syngeneic twins. Prior to transplantation, the donor and recipient are heavily typed to ensure that they are immunologically in close proximity. GVHD can also occur when there is an antigenic difference between the donor and recipient minor histocompatibility antigens. Therefore, GVHD may also occur between MHC-matched people. In addition, GVHD can also occur in surgical patients receiving directed transfusions, such as transfusions from HLA homozygous children to heterozygous parents. In certain embodiments, GVHD occurs when blood is infused into an immunocompromised patient (e.g., an organ transplant patient under high dose immunosuppressive agents, a child with primary immunodeficiency, or an HIV infected patient with AIDS).

There are two known forms of GVHD-acute and chronic GVHD. Acute GVHD may occur within the first 100 days after transplantation. Without wishing to be bound by a particular theory, it is believed that T-cells present in the donor's tissue and/or cells at the time of transplantation may attack the patient's skin, liver, stomach, and/or intestine. The earliest signs of acute GVHD can be rashes appearing on the hands, feet and face. In addition to blistering skin, patients with severe GVHD can also develop massive watery or bloody diarrhea with cramps due to attack of the stomach and intestines by donor T-cells. Jaundice (yellowing of the skin and eyes) is a common indication that GVHD disease involves the liver. The severity of acute GVHD disease can be assessed by the number of organs involved and the extent of symptoms.

Cases of acute GVHD can be divided into different stages according to clinical severity (see, e.g., Irani, J. et al, "Severe acute gastrointestinal graft versus host disease: emerging surgical dilemma in contemporary cancer care" (search access scientific grade-vs. host disease: an empirical clinical dilemma in clinical cancer care), Arch Surg.2008; 143(11): 1041-5). Stage 1 includes rashes on less than 25% of the body. Stage 2 includes a rash on more than 25% of the body with mild hepatic or gastrointestinal disorders. Stage 3 included redness of the skin and moderate liver, stomach and bowel problems similar to severe sunburn. Stage 4 included blistering, flaky skin and severe liver, stomach and bowel problems. Additionally or alternatively, acute GVHD can also be characterized by 5 clinical grades 0, I, II, III and IV. Generally, grade 0 is essentially asymptomatic for skin, liver, bowel or functional lesions. Grade I is considered mild with a stage 1 to 2 skin. Grade II is considered moderate and is characterized by having a phase 1 to 3 skin, a phase 1 liver and intestine and a phase 1 functional impairment. Grade III is considered severe and is characterized by stages 2 to 3 skin, 2-3 liver and intestine, and stage 2 functional damage. Grade IV is considered life threatening and is characterized by stage 2 to 4 skin, stage 2 to 4 liver and intestine, and stage 3 functional damage. Exemplary detailed staging and staging are further described in the examples section.

Chronic GVHD can occur after the first 100 days post-transplant. Without wishing to be bound by a particular theory, it is believed that chronic GVHD may be caused by T-cells produced by the implanted tissue and/or cells. The same organs and systems can be attacked as in acute GVHD, and in addition, chronic GVHD may be associated with connective tissue damage. Patients with chronic GVHD may experience skin problems that may include dry pruritic rashes, changes in skin color, and taut or taut skin. Partial alopecia or premature whitening may also occur. Similar to patients with acute GVHD, patients with chronic GVHD may exhibit jaundice as a sign of involvement of the liver. Chronic GVHD can also attack glands that secrete mucus, saliva, or other lubricants in the body. Patients with chronic GVHD may experience dryness or stinging in the eye due to impaired lacrimal glands. Salivary glands in the mouth may also be affected by chronic GVHD and in rare cases glands lubricating the oesophagus, resulting in swallowing and eating difficulties. Patients with chronic GVHD may experience a burning sensation in the mouth when using toothpaste or eating acidic foods. Chronic GVHD can attack glands that lubricate the stomach wall and intestinal tract, interfering with the body's ability to normally absorb nutrients. Symptoms may include heartburn, stomach ache and/or weight loss. Sometimes, patients with chronic GVHD may experience "contracture," i.e., tendon tightening, which makes it difficult to extend or contract the arms and legs. Chronic GVHD can also affect the lungs, causing wheezing, bronchitis, and/or pneumonia.

The incidence of GVHD can increase with increasing degree of mismatch between HLA antigens of the donor and recipient, increasing age of the donor, and increasing age of the patient. However, the disease may not be adequately diagnosed and reported.

As described above, GVHD can occur after a recipient receives a transplant of allogeneic cells, tissues or organs (e.g., cord blood, bone marrow, adult stem cells, embryonic stem cells, blood products, and/or solid organs).

Adsorbent and adsorbent preparation

The term "adsorbent" refers to any compound or material that can generally adsorb a substance to be adsorbed by a physicochemical bond between its surface and the substance. The adsorbent may be specific or non-specific. Preferred adsorbents for use in the present invention are pharmaceutical grade adsorbents, preferably suitable for pharmaceutical or veterinary applications in humans or animals.

Examples of adsorbents suitable for use in the present invention include, but are not limited to, activated carbon (also known as activated carbon); clays, including bentonite, kaolin, montmorillonite, attapulgite, halloysite, laponite, and the like; silicas, including colloidal silicas (e.g., silica)AS-40), mesoporous silica (MCM41), fumed silica, zeolites, and the like; talc; cholestyramine, and the like; polystyrene sulfonate and the like; mono-and poly-sulfonated resins; and other resins, e.g. for bacteriological examination e.g.And (3) resin.

Preferred adsorbents are pharmaceutical grade activated Carbons (e.g., from Chemviron, Cabot, Norit, Jacobi Carbons, Merck Millipore, Sigma Aldrich, Desotec, or other sources). In a particular embodiment, the adsorbent is activated carbon, more particularly with a height higher than 600m²A/g, in particular above 800m²A/g, in particular above 1000m²A/g, in particular above 1200m²A/g, in particular above 1400m²A/g, in particular above 1600m²G, even more particularly higher than 1800m²Activated carbon per g of specific surface area. The activated carbon may be of vegetable, mineral or synthetic origin, the surface of which is optionally modified by physical or chemical treatment. In a particular embodiment, the activated carbon is of plant origin. In a particular embodiment, the activated carbon is derived from peat. In a particular embodimentWherein the activated carbon is derived from coconut shells. In certain embodiments, the activated carbon is derived from different sources such as peat and coconut shell mixed together. In a particular embodiment, the activated carbon is characterized by a european molasses number preferably higher than 100, even more particularly higher than 200, even more particularly higher than 300, even more particularly higher than 400, even more particularly higher than 500, even more particularly higher than 600 (it should be noted that the european molasses number is inversely related to the north american molasses number). In a particular embodiment, the activated carbon has an antipyrine value (measured according to the european pharmacopoeia (EU pharmacopoeia)) higher than 10g/100g, even more particularly higher than 20g/100g, even more particularly higher than 30g/100g, even more particularly higher than 40g/100g, even more particularly higher than 50g/100g, even more particularly higher than 60g/100 g. In a particular embodiment, the activated carbon is characterized by having a density of between 0.05 and 0.8, even more particularly between 0.1 and 0.6, even more particularly between 0.15 and 0.5, even more particularly between 0.2 and 0.4.

The amount of adsorbent used in the process of the invention may vary with the host/material to be treated and the total capacity, adsorption capacity and selectivity of the adsorbent. Generally, the amount of adsorbent is an amount sufficient to prevent the deleterious effects of substances such as antibiotics on the intestinal microflora known as "dysbiosis" or destruction of the intestinal microflora. In particular, the amount of adsorbent is an amount sufficient to prevent or delay the onset or reduce the severity of acute and/or chronic GVHD.

The adsorbents used in the present invention may be formulated into compositions, e.g., pharmaceutical compositions, which may include pharmaceutically acceptable excipients, carriers, and/or additives. Such compositions include formulations for oral delivery, rectal delivery, topical administration, mucosal administration, inhalation, and the like. In particular embodiments, the adsorbent is formulated as a pharmaceutical composition suitable for administration to a human or animal. More preferably, the adsorbent is formulated as an oral formulation suitable for releasing the adsorbent in the intestine or upon contact with intestinal bacteria, in particular in the gastrointestinal tract, more in particular in the lower part of the intestine, i.e. the terminal ileum, caecum and/or colon.

The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. Examples of formulations suitable for intestinal delivery of adsorbents have been described in WO2006/122835 and WO 2007/132022. In another embodiment, the adsorbent is formulated in a core. Preferably, the amount of adsorbent is between about 60% to about 100%, more preferably between about 70% to about 98%, more preferably between about 75% to about 95%, more preferably between about 80% to about 90% of the total weight of the core. In a preferred embodiment, the adsorbent is formulated with carrageenan, preferably in the form of pellets, as proposed in WO 2011/104275. Such a formulation may form a core. Such a core may be covered with a coating layer such that the adsorbent is released in the lower part of the intestine, i.e. in the terminal ileum, caecum and/or colon. Alternatively, the plurality of cores may be contained or embedded in a dosage unit form adapted to release its contents in the lower part of the intestine, i.e. in the terminal ileum, caecum and/or colon, e.g. a capsule having a shell adapted to release its contents in the lower part of the intestine. In another embodiment, the pellets may be contained in a capsule, which itself is contained in a coated capsule. In another embodiment, the pellet may be contained or embedded in a multi-unit particle system.

Carrageenans are a naturally occurring linear sulfated polysaccharide extracted from red seaweed. Carrageenan is a high molecular weight polysaccharide composed of repeating sulfated and nonsulfated galactose and 3, 6-anhydrogalactose (3,6-AG) units. The units are linked by alternating alpha 1-3 and beta 1-4 glycosidic linkages. Three basic types of carrageenan are commercially available, namely kappa, iota and lambda carrageenan, which differ by the number and location of sulfate groups on the galactose units. The carrageenan used in the present invention may be selected from kappa, iota and lambda carrageenan and mixtures thereof. In one aspect of this embodiment, the adsorbent is mixed with kappa-carrageenan. In a particular embodiment, the mixture comprises activated carbon and kappa-carrageenan. Preferably, the amount of carrageenan is between about 5% and about 25%, more preferably between about 10% and about 20% of the total weight of the adsorbent and carrageenan. In other particular embodiments, the amount of adsorbent (particularly activated carbon) in the mixture is between about 95% to about 75%, more preferably between about 90% to about 80% of the total weight of the adsorbent and carrageenan. According to a particular embodiment of the invention, the amount of carrageenan is about 15% of the total weight of the adsorbent and carrageenan. For example, the mixture may contain 85% adsorbent and 15% carrageenan.

According to a particular embodiment of the present invention, a mixture of activated carbon and carrageenan, in particular kappa-carrageenan, having the weight ratio specified above is provided.

The core (or pellet) may be produced by any suitable means known to the skilled person. In particular, the granulation technique is modified to adapt to the production of the core. For example, the core may be obtained as follows: the adsorbent is mixed with carrageenan in the ratios indicated above, a solvent such as water is added for wet granulation, followed by extrusion, optionally followed by spheronization or spheronization with a roller knife or by a one-pot spheronization. Any remaining water can be removed, for example, by drying the resulting pellets using conventional techniques.

In one embodiment, the core or pellet has an average particle size in the range of from 50 μm to 6000 μm, in particular from 100 μm to 5000 μm, in particular from 150 μm to 4000 μm, in particular from 250 to 3000 μm, in particular from 250 to 1000 μm, in particular from 300 to 3000 μm (e.g. from 500 to 3000 μm), in particular from 300 to 1000 μm, in particular from 500 to 700 μm.

The core composition may also comprise conventional excipients such as anti-adherents, binders, fillers, diluents, flavouring agents, colouring agents, lubricants, glidants, preservatives, adsorbents and/or sweeteners. The amount of such excipients may vary, but is typically in the range of 0.1 to 50% by weight of the pellet.

As discussed above, preferred formulations of the present invention comprise a core containing the adsorbent and possibly supplemented with carrageenan, said core being covered by a coating layer such that the adsorbent is released in the lower part of the intestine, i.e. in the terminal ileum, caecum and/or colon.

In this regard, in a preferred embodiment, the adsorbent is used as a formulation comprising:

-a core containing said adsorbent, and

-an outer coating layer formed around the core such that the adsorbent is released from the formulation in the lower part of the intestine.

In a preferred embodiment, the adsorbent is used as a formulation comprising:

-a core comprising said adsorbent and carrageenan, and

-an outer coating layer formed around the core such that the adsorbent is released from the formulation in the lower part of the intestine.

Examples of suitable coatings include pH-dependent enteric polymers, azo-based polymers, disulfide polymers and polysaccharides, in particular amylose, pectin (e.g. pectin coupled with divalent cations such as calcium or zinc pectin), chondroitin sulphate and guar gum. Representative pH-dependent enteric polymers include Cellulose Acetate Trimellitate (CAT), Cellulose Acetate Phthalate (CAP), acrylic polymers, methacrylic polymers, anionic copolymers based on methyl acrylate, methyl methacrylate and methacrylic acid, hydroxypropyl methylcellulose phthalate (HPMCP), hydroxypropyl methylcellulose acetate succinate (HPMCAS), copolymers of methacrylic acid and ethyl acrylate, in a molar ratio of 1:1 copolymer of methacrylic acid and methyl methacrylate, in a molar ratio of 1:2, a copolymer of methacrylic acid and methyl methacrylate, polyethylene acetate phthalate (PVAP), and shellac resin. Particularly preferred polymers include shellac, anionic copolymers based on methyl acrylate, methyl methacrylate and methacrylic acid, for example in a molar ratio of 7:3:1 (poly (methyl acrylate-co-methyl methacrylate-co-methacrylic acid), and a molar ratio of 1:2 of methacrylic acid and methyl methacrylate. Ideally, the polymer dissolves at a pH equal to 6.0 and higher, preferably 6.5 and higher. Suitable coatings may also be obtained by mixing the above mentioned polymers and copolymers. In another embodiment, a suitable coating is a time-dependent coating or is based on a mixture of a time-dependent polymer such as ethylcellulose polymer with sodium alginate.

In a particular embodiment, the formulation comprises a further intermediate coating between the core and the outer pH-dependent layer. The intermediate coating may be formed from a variety of different polymers, including pH-dependent polymers, pH-independent water-soluble polymers, pH-independent insoluble polymers, and mixtures thereof. Examples of such pH-dependent polymers include shellac type polymers, anionic copolymers based on methyl acrylate, methyl methacrylate and methacrylic acid, copolymers of methacrylic acid and ethyl acrylate, hydroxypropyl methylcellulose phthalate (HPMCP) and hydroxypropyl methylcellulose acetate succinate (HPMCAS). Examples of pH independent water soluble polymers include PVP or high molecular weight cellulose polymers such as hydroxypropyl methylcellulose (HPMC) or hydroxypropyl cellulose (HPC). Examples of pH independent insoluble polymers include ethyl cellulose polymers or copolymers of ethyl acrylate and methyl methacrylate.

In a particular embodiment, the invention uses a formulation comprising:

a core comprising a mixture of adsorbent (preferably activated carbon) and carrageenan (preferably kappa-carrageenan),

an intermediate coating selected from HPMC, ethylcellulose and copolymers of methacrylic acid and ethyl acrylate, for exampleCopolymers of L30D-55 with ethyl acrylate and methyl methacrylate, for exampleA mixture of NE30D (e.g. a mixture in a weight ratio of 1:9 to 9:1, preferably 2:8 to 3: 7), and

an outer layer made of an anionic copolymer based on methyl acrylate, methyl methacrylate and methacrylic acid, such as 7:3:1 poly (methyl acrylate-co-methyl methacrylate-co-methacrylic acid), for exampleFS 30D.

In a particular embodiment, the formulation comprises a core comprising about 85% activated carbon and about 15% kappa-carrageenan and a coating having an anionic copolymer based on methyl acrylate, methyl methacrylate and methacrylic acid (e.g., 7:3:1 poly (methyl acrylate-co-methyl methacrylate-co-methacrylic acid), e.g., methyl acrylate-co-methyl methacrylate-co-methacrylic acid)FS30D, Evonik, Darmstadt, Germany) or copolymers of methacrylic acid and ethyl acrylate (e.g.L30D55, Evonik, Darmstadt, Germany).

In another embodiment, the adsorbent is formulated as a composition as disclosed in WO2014044794 comprising:

(a) a core comprising activated carbon;

(b) a first layer surrounding the core, the first layer comprising an insoluble semi-permeable material; and

(c) a second layer surrounding the first layer that dissolves at a predetermined pH or at a predetermined location in the gastrointestinal tract.

In a variation of this embodiment, the core is activated carbon. In another variation, the activated carbon is grit blasted or deflashed. In yet another variation, the activated carbon has a particle size of 0.02 to 5.0mm, such as 0.6 to 1.2 mm. In another variation, the insoluble semipermeable material comprises one or more of ethylcellulose, glyceryl monostearate, cellulose butyrate acetate, dimeric lactic acid, polyvinyl chloride, and poly (meth) acrylate polymers such as Eudragit RL 100, Eudragit RL PO, Eudragit RL 30D, Eudragit RL 12.5, Eudragit RS 100, Eudragit RS PO, Eudragit RS 30D, Eudragit RS 12.5, and Eudragit NE30D, Eudragit HE 40D. In another variation, the first layer further comprises a water soluble material, wherein the first layer further comprises a water soluble material comprising Hydroxypropylmethylcellulose (HPMC). In certain embodiments, the water-soluble material may be mixed with the insoluble semipermeable material, and/or may comprise the insoluble semipermeable material in an amount of 0.1 to 30% by weight, for example in an amount of 2 to 25% by weight. In another particular variant, the first layer allows the gradual diffusion of molecules through the semi-permeable membrane towards the core, coming into contact with the activated carbon. In yet another variation, the second layer comprises a material that dissolves at pH 5 to pH 7. In certain variations, the second layer is an enteric layer comprising a material that remains substantially intact at a pH of 1 to 4.9 but rapidly decomposes at a pH of 5 to 7. In one variation, the second layer comprises a pH-sensitive polymer. Representative second layers include layers selected from the group consisting of: hydroxypropyl methylcellulose acetate succinate, Cellulose Acetate Trimellitate (CAT), Cellulose Acetate Phthalate (CAP), anionic copolymers based on methyl acrylate, methyl methacrylate and methacrylic acid, hydroxypropyl methylcellulose phthalate (HP CP), hydroxypropyl methylcellulose acetate succinate (HPMCAS), copolymers of methacrylic acid and ethyl acrylate, copolymers of methacrylic acid and methyl methacrylate (1:1 molar ratio), copolymers of methacrylic acid and methyl methacrylate (1:2 molar ratio), polyethylene acetate phthalate (PVAP) and shellac resin. In another particular variation of this embodiment, the activated carbon is the only active pharmaceutical ingredient. In yet another variation, the composition comprises:

(a) a core comprising activated carbon;

(b) a first layer surrounding the core, the first layer comprising an insoluble semipermeable material in the form of ethylcellulose, and optionally further comprising a water-soluble material comprising Hydroxypropylmethylcellulose (HPMC); and

(c) a second layer comprising hydroxypropyl methylcellulose acetate succinate (HPMCAS).

In another variation, the adsorbent is activated carbon formulated in a composition comprising:

(a) a core, which is activated carbon;

(b) a first layer surrounding said core, said first layer comprising a water-insoluble semipermeable material and further comprising a water-soluble material comprising hydroxypropylmethylcellulose in an amount of 2-25% by weight of said insoluble semipermeable material; and

(c) a second layer surrounding the first layer, which dissolves at a pH of 5 to 7.

Antibiotic

The term "antibiotic" refers to any compound having activity against bacteria. Antibiotics that may be eliminated or inactivated, thanks to the present invention, include but are not limited to:

- β -lactams comprising:

penicillins (e.g., penicillin G, penicillin V, ampicillin, amoxicillin, bacampicillin, carbenicillin, ticarcillin, azlocillin, mezlocillin, piperacillin, etc.),

penicillinase-resistant penicillins (e.g., methicillin, oxacillin, cloxacillin, dicloxacillin, nafcillin, etc.),

cephalosporins, for example: first generation cephalosporins (e.g. cefadroxil, cephalexin, cephradine, cephalothin, cefapirin, cefazolin, etc.); second generation cephalosporins (e.g., cefaclor, cefamandole, cefonicid, cefoxitin, cefotetan, cefuroxime axetil, cefmetazole, cefprozil, chlorocefcapene, cefradide, etc.); third-generation cephalosporins (e.g., cefepime, cefoperazone, cefotaxime, ceftizoxime, ceftriaxone, ceftazidime, cefixime, cefpodoxime, ceftibuten, etc.); fourth generation cephalosporins (e.g. cefixdine, cefepime, cefozopran, cefpirome, cefquinome, etc.); fifth and other generation cephalosporins (e.g. cefepime, ceftaroline, ceftizolidine, etc.),

carbapenems (e.g. imipenem, meropenem, ertapenem, doripenem, etc.),

monocyclic lactams (e.g. aztreonam and the like),

quinolones (e.g. nalidixic acid) and fluoroquinolones (e.g. cinoxacin, ciprofloxacin, moxifloxacin, levofloxacin, ofloxacin, gatifloxacin, gemifloxacin, norfloxacin etc.),

sulfonamides (e.g. sulfanilamide, sulfadiazine, sulfamethoxazole, sulfisoxazole, sulfacetamide, sulfamethoxydiazine, etc.),

aminoglycosides (e.g. streptomycin, gentamicin, tobramycin, amikacin, netilmicin, kanamycin, neomycin B, C and E, spectinomycin, puromycin, gentamicin and the like),

tetracyclines (e.g. tetracycline, chlortetracycline, oxytetracycline, methacycline, doxycycline, minocycline, tigecycline, elacycline, etc.),

macrolides (e.g. erythromycin, azithromycin, clarithromycin, fidaxomycin, telithromycin, josamycin, oleandomycin, spiramycin, tylosin, roxithromycin, quinomycin, solithromycin, etc.),

glycopeptides (e.g. vancomycin, oritavancin, telavancin, teicoplanin, dalbavancin, ramoplanin, etc.),

oxazolidinones (e.g. linezolid, epsiprazole, tedizolid, ralazolide, cycloserine, etc.),

chloramphenicol (e.g., chloramphenicol, thiamphenicol, etc.),

lincosamides (e.g. clindamycin, lincomycin, etc.),

streptogramins (e.g. pristinamycin, quinupristin/dalfopristin, virginiamycin, etc.),

polymyxins (e.g. polymyxin A, B, C, D, E1 (colistin A) or E2, colistin B or C etc.),

diaminopyrimidines (e.g., trimethoprim, pyrazinamide, etc., commonly used in combination with sulfamethoxazole),

sulfones (e.g. dapsone, sodium aclidinesulfonate, etc.),

-a source of para-aminobenzoic acid,

-a bacitracin which is capable of inhibiting the growth of bacillus,

-a source of isoniazid,

rifampins (e.g. rifampin, rifabutin, rifapentine, rifalazil, rimamixin, etc.),

-a source of-ethyl-amine-butanol,

-a source of N-methyl-aminothioisonicotinamine,

-a capreomycin,

-clofazimine, and

-any other antibacterial agent.

The term "antibiotic" also covers combinations of antibiotics.

Antibiotic inactivating enzymes and enzyme preparations

In certain embodiments, the invention uses antibiotics to inactivate enzymes. In the context of the present invention, an "antibiotic-inactivating enzyme" is an enzyme capable of hydrolyzing or inactivating an antibiotic, thereby rendering the antibiotic biologically inactive. For example, an antibiotic-inactivating enzyme may substantially increase the Minimum Inhibitory Concentration (MIC) of an antibiotic as compared to the MIC of the antibiotic obtained in the absence of the antibiotic-inactivating enzyme. According to the invention, antibiotic inactivation is complete if the growth of bacteria sensitive to a certain concentration of a given antibiotic in the presence of said concentration of antibiotic after treatment with said inactivating enzyme is identical to the growth in the absence of said antibiotic. Another definition of total inactivation is that the MIC of an antibiotic for a susceptible bacterium increases by at least 2 orders of magnitude after treatment with the inactivating enzyme.

The antibiotic-inactivating enzyme used according to the present invention may be natural, chemically modified, genetically engineered or synthetic.

Antibiotic-inactivating enzymes also include functional variants of parent antibiotic-inactivating enzymes, such as functional variants of beta-lactamases, erythromycin esterases, and ketoreductases. In the context of the present invention, a "functional variant" of an enzyme is an enzyme derived from a parent enzyme, which has the same type of catalytic activity (e.g., a beta-lactamase variant is an enzyme having beta-lactamase activity), but has a different amino acid sequence. Such functional variants may have at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, at least 99.9% identity to the parent enzyme. Such functional variants may also have a specific activity for a given antibiotic, e.g., in the case of a beta-lactamase, of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 105%, 110%, 115%, 120%, 125%, 130%, 135%, 140%, 145%, 150%, 160%, 170%, 180%, 190%, 200%, 220%, 240%, 260%, 280%, 300%, 350%, 400%, 500%, 600%, 700%, 800%, or even at least 1600% relative to the specific activity of the parent antibiotic-inactivating enzyme.

Representative antibiotic-inactivating enzymes useful in the practice of the present invention include, but are not limited to, enzymes that inactivate beta-lactam antibiotics (e.g., beta-lactamases), fluoroquinolone-inactivating enzymes (e.g., aminoglycoside N-acetyltransferases), macrolide-inactivating enzymes (e.g., erythromycin esterases or erythromycin phosphotransferases), tetracycline-inactivating enzymes (e.g., NADPH-dependent tetracycline oxidoreductases), or lincosamide-inactivating enzymes (e.g., lincomycin nucleotidyl transferases).

Beta-lactamases are enzymes (EC 3.5.2.6) having beta-lactamase activity, i.e. enzymes which catalyze the irreversible hydrolysis of the amide bond of the beta-lactam ring present in compounds such as beta-lactam antibiotics (e.g. penicillins, cephalosporins, carbapenems, penem sulfones) to produce a hydrolyzed molecule without its antibacterial activity. Such enzymes are well known to those skilled in the art (Wang et al, 1999, Curr Opin Chem biol.3(5), 614-22; frere, J.M.1995, Mol Microbiol.16(3): 385-95).

In particular embodiments, the beta-lactamase is a serine beta-lactamase or a zinc-dependent beta-lactamase, also known as a metallo beta-lactamase. In another embodiment, the beta-lactamase is selected from the group consisting of class a, class B, class C, and class D beta-lactamases. In other specific embodiments, the beta-lactamase is selected from group 1, group 2, group 3, and group 4 beta-lactamases (Bush et al, antibiotic. Agents Chemother,39: 1211). In certain embodiments, the beta-lactamase is one or more of P1A, P3A, or P4A and derivatives thereof consisting of derivatives of beta-lactamase from Bacillus licheniformis (Bacillus licheniformis) 749/C or P2A and derivatives thereof of metallo beta-lactamase from Bacillus cereus. Further, the beta-lactamase may be a broad spectrum beta-lactamase (ESBL), optionally selected from TEM, SHV, CTX-M, OXA, PER, VEB, GES, and IBC beta-lactamase. Furthermore, the beta-lactamase may be an inhibitor resistant beta-lactamase, optionally selected from the group consisting of AmpC type beta-lactamases, carbapenemases such as, but not limited to, IMP type carbapenemases (metallo beta-lactamases), VIM (veronian integrant encoded metallo beta-lactamase) carbapenemases, OXA (oxacillinase) group beta-lactamases, KPC (klebsiella pneumoniae (k. pneumonia) carbapenemases), CMY (C-class), SME, IMI, NMC and CcrA, and NDM (new delrin metallo beta-lactamases such as NDM-1) beta-lactamases.

In certain embodiments, the beta-lactamase is VIM (a metallo beta-lactamase encoded by the veraron integrant). Illustrative VIM enzymes include, but are not limited to, VIM-1, VIM-2, VIM-3, VIM-4, and VIM-19. Additional VIM enzymes are described, for example, in Queenan et al, (2007) Clin. Microbiol. Rev.20(3): 440-458. In other particular embodiments, the beta-lactamase is VIM-2 or a variant thereof. Such beta-lactamases are disclosed in PCT/EP2017/053985, PCT/EP2017/053986 and EP 17198414. In particular instances, the present invention relates to the use of any particular embodiment disclosed in PCT/EP2017/053985, PCT/EP2017/053986 and EP17198414, including any particular variant VIM-2 disclosed therein. In certain embodiments, the antibiotic-inactivating enzyme is, for example, an antibiotic-inactivating enzyme as set forth in SEQ ID NO: VIM-2 as shown in 1. In another specific embodiment, the antibiotic-inactivating enzyme is a polypeptide having the amino acid sequence as set forth in SEQ ID NO: 2 to 46, or a VIM-2 functional variant of the amino acid sequence set forth in seq id no. In certain embodiments, the VIM-2 functional variant has a sequence comprising SEQ ID NO: 29. SEQ ID NO: 31. SEQ ID NO: 34 or SEQ ID NO: 36 or a sequence consisting thereof.

In another embodiment, the beta-lactamase is a beta-lactamase from Bacillus licheniformis (Bacillus licheniformis) 749/C or a variant thereof, such as P1A, P3A (also known as "ribaxamase"), or P4A. P1A has the amino acid sequence of SEQ ID NO: 47.

In certain embodiments, the beta-lactamase is a metallobeta-lactamase (also referred to as P2A) from Bacillus cereus (Bacillus cereus) or a functional variant thereof, as described in, for example, WO 2007147945. In certain embodiments, the P2A enzyme has the amino acid sequence of SEQ ID NO: 48. The functional variant of the P2A enzyme may be identical to SEQ ID NO: 48 have at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, at least about 98%, at least about 99%, or 100% identity.

In certain embodiments, the beta-lactamase is P3A or a functional variant thereof as described, for example, in WO 2011148041. In certain embodiments, the P3A enzyme has the amino acid sequence of SEQ ID NO: 49 (mature form of the enzyme) or SEQ ID NO: 50 (a form of the enzyme comprising a signal peptide 31 amino acids long). The functional variant of the P3A enzyme may be identical to SEQ ID NO: 49 or SEQ ID NO: 50 have at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, at least about 98%, at least about 99%, or 100% identity. In certain embodiments, the beta-lactamase comprises a sequence identical to SEQ ID NO: 49 and characterized in that it has a hydrophilic amino acid residue other than aspartic acid (D) at a position corresponding to position 276 of the classification according to Ambler, and in that the hydrophilic amino acid is selected from the group consisting of arginine (R), histidine (H), lysine (K), asparagine (N), glutamine (Q), serine (S) and threonine (T). In other particular embodiments, the beta-lactamase comprises an amino acid sequence identical to SEQ ID NO: 49, and characterized in that it has asparagine (N) at the position corresponding to position 276 of the classification according to Ambler. In yet another embodiment, the beta-lactamase has an amino acid sequence of SEQ ID NO: 49, wherein the amino acid residue at the position corresponding to position 276 according to the classification by Ambler is asparagine (N).

In another embodiment, the beta-lactamase is P4A as described in, e.g., WO2015/161243, or a functional variant thereof. In certain embodiments, the P4A enzyme has the amino acid sequence of SEQ ID NO: 79 or SEQ ID NO: 80 in a sequence of seq id no. The functional variant of the P4A enzyme may be identical to SEQ ID NO: 79 or SEQ ID NO: 80 have at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, at least about 98%, at least about 99%, or 100% identity.

In certain embodiments, the beta-lactamase is Klebsiella pneumoniae (Klebsiella pneumoniae) carbapenemase (KPC). Illustrative KPCs include, but are not limited to KPC-1/2(SEQ ID NO: 51), KPC-3(SEQ ID NO: 52), KPC-4(SEQ ID NO: 53), KPC-5(SEQ ID NO: 54), KPC-6(SEQ ID NO: 55), KPC-7(SEQ ID NO: 56), KPC-8(SEQ ID NO: 57), KPC-9(SEQ ID NO: 58), KPC-10(SEQ ID NO: 59), KPC-11(SEQ ID NO: 60), KPC-12(SEQ ID NO: 61), KPC-13(SEQ ID NO: 62), KPC-14(SEQ ID NO: 63), KPC-15(SEQ ID NO: 64), and KPC-17(SEQ ID NO: 65). In one embodiment, the beta-lactamase is KPC-1/2. In one embodiment, the beta-lactamase is KPC-3. A functional variant of a KPC enzyme may be compared to SEQ ID NO: 51 to SEQ ID NO: 65 have at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, at least about 98%, at least about 99%, or 100% identity.

In another embodiment, the beta-lactamase is a neodrime beta-lactamase (NDM). Illustrative NDMs include, but are not limited to NDM-1(SEQ ID NO: 66), NDM-2(SEQ ID NO: 67), NDM-3(SEQ ID NO: 68), NDM-4(SEQ ID NO: 69), NDM-5(SEQ ID NO: 70), NDM-6(SEQ ID NO: 71), NDM-7(SEQ ID NO: 72), NDM-8(SEQ ID NO: 73), NDM-9(SEQ ID NO: 74), NDM-10(SEQ ID NO: 75), NDM-11(SEQ ID NO: 76), NDM-12(SEQ ID NO: 77), and NDM-13(SEQ ID NO: 78). In one embodiment, the beta-lactamase is NDM-1. In one embodiment, the broad spectrum carbapenemase is NDM-4. Functional variants of NDM enzymes may be compared to SEQ ID NO: 66 to SEQ ID NO: 78, has at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, at least about 98%, at least about 99%, or 100% identity.

In certain embodiments, the beta-lactamase is an IMP type carbapenemase. Illustrative IMP-type enzymes include, but are not limited to, IMP-1, IMP-4, IMP-8, IMP-11, IMP-43, and IMP-44. Additional IMP-type enzymes are described, for example, in Queenan et al, (2007) Clin. Microbiol. Rev.20(3): 440-458.

In certain embodiments, the beta-lactamase is from an OXA (oxacillin enzyme) group of beta-lactamases. Illustrative OXA β -lactamases include, but are not limited to, OXA-23, OXA-24, OXA-27, OXA-40, OXA-48, OXA-49, OXA-50, OXA-51, OXA-58, OXA-64, OXA-71, and OXA-181. Additional beta-lactamases of the OXA type are described, for example, in Walther-Rasmussen et al, Journal of analytical chemistry (2006),57:373-383 and Queenan et al, (2007) Clin. Microbiol. Rev.20(3): 440-458.

In certain embodiments, the beta-lactamase is a CMY (carbapenemase class C) enzyme. An illustrative CMY enzyme having carbapenemase activity is CMY-10, as described, for example, in Lee et al, (2006) Research Journal of Microbiology (1): 1-22.

In certain embodiments, the beta-lactamase is an SME enzyme (representing serratia marcescens). Illustrative SME enzymes include, but are not limited to, SME-1, SME-2 or SME-3, as described, for example, in Queenan et al, (2007) Clin. Microbiol. Rev.20(3): 440-458.

In certain embodiments, the beta-lactamase is an IMI enzyme (beta-lactamase that hydrolyzes imipenem). Illustrative IMI enzymes include, but are not limited to IMI-1 or IMI-2, as described, for example, in Queenan et al, (2007) Clin. Microbiol. Rev.20(3): 440-458.

In certain embodiments, the beta-lactamase is an NMC enzyme (a carbapenemase that is not a metalloenzyme). An illustrative NMC enzyme is NMC-A, as described, for example, in Queenan et al, (2007) Clin. Microbiol. Rev.20(3): 440-458.

In certain embodiments, the beta-lactamase is a GES enzyme (guiana extended spectrum). Illustrative GES enzymes include, but are not limited to, GE-2, GES-4, GES-5, GES-6, GES-7, GES-8, GES-9, GES-11, GES-14, and GES-18, as described, for example, in Queenan et al, (2007) Clin.Microbiol.Rev.20(3): 440-.

In certain embodiments, the beta-lactamase is ccra (cfia) metallo beta-lactamase from Bacteroides fragilis (Bacteroides fragiliss).

In certain embodiments, the beta-lactamase is the SFC-1 enzyme from Serratia fonticola (Serratia fonticola) or the SHV-38 enzyme from Klebsiella pneumoniae (Klebsiella pneumoniae), as described, for example, in Walther-Rasmussen et al, (2007) Journal of Antimicrobial Chemotherapy,60: 470-482.

In another embodiment, the antibiotic-inactivating enzyme is an erythromycin esterase. Erythromycin esterase (EC number 3.1.1) refers to a class of enzymes that catalyze the inactivation of erythromycin and other macrolide antibiotics. These enzymes hydrolyze the lactone rings of macrolides such as erythromycin and oleandomycin as explained in Barthelemy et al, 1984, J.Antibiott.37, 1692-1696. Known erythromycin esterases are of bacterial origin. They are produced, for example, by Escherichia coli (Escherichia coli), Halobacterium salinarum, Gramedla forsetii, Achromobacter denitrificans (Achromobacter denificans) or Rhodococcus sp. In a particular embodiment, the erythromycin esterase is one of the enzymes normally produced by members of the Enterobacteriaceae family that are highly resistant to erythromycin, as described in Arthur et al, 1987, Antimicrob. Agents Chemother.31(3), 404-. Two erythromycin esterases from E.coli have been recorded under the reference names EreA and EreB, the use of both of which is envisaged in the present invention. In a particular embodiment of the invention, the erythromycin esterase is an EreB erythromycin esterase from E.coli (see Arthur et al, 1986, Nucleic Acids Res 14(12), 4987-.

In another embodiment, the antibiotic-inactivating enzyme is a ketoreductase. Ketoreductase (KRED) or carbonyl reductase-like (EC 1.1.1.184) enzymes are useful for the synthesis of optically active alcohols from the corresponding prochiral ketone substrates. KREDs typically convert a ketone substrate to the corresponding alcohol product, but may also catalyze the reverse reaction, oxidizing the alcohol substrate to the corresponding ketone/aldehyde product.

In another embodiment, the antibiotic inactivating enzyme is a hybrid protein molecule. Representative hybrid protein molecules are those disclosed in U.S. patent application 20170354706. Such hybrid protein molecules may comprise two enzymes bonded together, capable of inactivating at least one antibiotic. In particular embodiments, these enzymes are combined into a single-chain protein. The two enzymes may both be from the same class, or each from a different class. For example, the two enzymes may be beta-lactamases, or selected from the class of beta-lactamases, aminoglycoside-inactivating enzymes, fluoroquinolone-inactivating enzymes, lincosamide-inactivating enzymes, macrolide-inactivating enzymes, or tetracycline-inactivating enzymes. In particular embodiments, each enzyme in the hybrid protein molecule inactivates a different antibiotic. In another embodiment, the hybrid protein molecule comprises two enzymes capable of inactivating antibiotics belonging to the same class. In certain embodiments, the sequence of at least one component enzyme in the hybrid protein is identical to SEQ ID NO: 81 to SEQ ID NO: 87 have at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8% or at least 99.9% sequence homology. In other particular embodiments, the sequence of at least one component enzyme in the hybrid protein has a sequence consisting of SEQ ID NO: 81. SEQ ID NO: 82. SEQ ID NO: 83. SEQ ID NO: 84. SEQ ID NO: 85. SEQ ID NO: 86 or SEQ ID NO: 87, respectively. In other particular embodiments, the hybrid protein molecule has a heavy chain comprising a sequence selected from SEQ ID NOs: 88 to 90 or an amino acid sequence consisting thereof.

In another embodiment, the enzyme, whether biologically or synthetically produced, may be further modified enzymatically and/or chemically to enhance its activity, stability, solubility, or any other beneficial characteristic. One example of such a modification is attachment of polyethylene glycol to a surface amino group or pegylation.

In particular embodiments, the antibiotic-inactivating enzyme is formulated in a formulation suitable for release of the enzyme in a desired portion of the intestine. In a particular embodiment, the desired portion of the intestine is the lower part of the intestine, such as the ileum, caecum or colon. In another specific embodiment, the desired portion of the intestine is the upper portion of the intestine, such as the duodenum or jejunum. In a particular embodiment, the formulation comprises pellets of the enzyme coated with an enteric coating (e.g., with an enteric coating that dissolves at a pH greater than or equal to 7.0). In another particular embodiment, the formulation comprises an enterically coated enzyme pellet (e.g., having an enteric coating that dissolves at a pH greater than or equal to 5.5 or a pH greater than or equal to 7.0) within an enterically coated capsule (e.g., having an enteric coating that dissolves at a pH greater than or equal to 5.5 or a pH greater than or equal to 7.0). In another specific embodiment, the formulation comprises enteric coated pellets in an uncoated capsule. The choice of the formulation may depend on the route of administration of the antibiotic to the subject. For example, in the case of parenteral administration of the antibiotic, preparations which release the antibiotic-inactivating enzyme in the upper or lower part of the intestine may be considered. In the case of oral administration of the antibiotic, the enzyme preparation is preferably released in the lower part of the intestine at a location where inactivation of the antibiotic by the enzyme is no longer able to interfere with the desired absorption of the antibiotic in the small intestine, in order to benefit from the positive effect of the antibiotic.

Application method

The present invention relates to compositions and methods for treating or preventing or delaying GVHD or reducing the severity of GVHD based on the use of substances suitable for inactivating dysbiosis-causing agents.

In particular, the invention is useful for preventing the destruction of the microbial flora and preventing or delaying the onset or reducing the severity of GVHD in patients receiving allogeneic hematopoietic stem cell transplantation.

In certain cases, the substances according to the invention are used in a subject to whom, to whom or to whom an agent may be administered that may disturb the intestinal microbial flora of the subject is administered. Thanks to the present invention, the harmful effects of such agents can be prevented. The present invention therefore relates to a method of reducing the potentially harmful effects of an agent on the gut microflora of a subject who is or can be the recipient of an immunocompetent transplant, said method comprising administering to said subject an effective amount of a substance according to the invention suitable for inactivating an agent causing dysbiosis.

The agent that causes dysbiosis may be an agent that is administered to treat a pathological condition in the subject. In fact, certain agents may be administered in order to treat diseases, but may have a detrimental effect on the intestinal microflora when they reach the lower part of the intestine. The subject will still receive the agent to benefit from its desired effect, but on the other hand, should provide a solution that avoids its secondary effects. Illustrative agents that have this behavior include antibiotics. As provided above, antibiotics can be administered to a subject in order to treat a bacterial infection. However, since antibiotics can be designed to affect bacterial growth or survival, they threaten the intestinal microflora balance when they reach the lower part of the intestine and may cause dysbiosis. Other illustrative agents that may cause dysbiosis include, but are not limited to:

chemotherapeutic agents, such as taxanes (e.g. docetaxel, paclitaxel), anthracyclines (e.g. doxorubicin), topoisomerase inhibitors (e.g. etoposide), antimetabolites (e.g. methotrexate, cytarabine, 5-fluorouracil, gemcitabine), alkylating agents (e.g. melphalan), kinase inhibitors (e.g. erlotinib),

antifungal agents, such as voriconazole, amphotericin B liposomes, posaconazole,

antiviral agents, such as acyclovir, metisazone,

anti-inflammatory agents, such as aspirin, ibuprofen.

Thus, in a further aspect of the invention, the substances according to the invention are administered to a subject suffering from cancer and being, to be or have been treated with a drug substance causing dysbiosis, such as an antibiotic.

The adsorbent or antibiotic inactivating enzyme (if appropriate since the drug substance causing dysbiosis is an antibiotic) may even be administered to the subject long before transplantation. For example, the subject may have been selected as a transplant recipient, but the treatment cannot begin days, weeks, months, or years ago. In this case, if the subject suffers from a disease between these events that requires treatment with an agent causing dysbiosis, such as an antibiotic, it would be advantageous to prevent dysbiosis of the intestinal microflora by administering the adsorbents or antibiotic inactivating enzymes provided herein. Likewise, the adsorbent or antibiotic-inactivating enzyme may be administered to the subject even a long time after the transplantation day. In particular, rejection of the subject's transplant by the host may unfortunately occur. In this case, stopping systemic administration of the adsorbent or antibiotic-inactivating enzyme when the subject receives a drug substance causing dysbiosis, such as an antibiotic, may seriously impair the efficacy of future transplantation.

In certain embodiments, the adsorbent or antibiotic-inactivating enzyme is administered to the subject at about the same time as the agent causing the dysbiosis, e.g., an antibiotic. By "substantially simultaneously" is meant that the adsorbent or antibiotic inactivating enzyme is administered shortly before, simultaneously with and/or shortly after, preferably shortly before, the administration of the dysbiosis causing agent, in particular antibiotic. In a particular embodiment, the adsorbent or antibiotic-inactivating enzyme is administered within less than 30 minutes before or after the dysbiosis-causing agent, in particular antibiotic, is administered, preferably within less than 20 minutes, less than 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 minutes or less than 1 minute before or after the dysbiosis-causing agent, in particular antibiotic, is administered. In other particular embodiments, the adsorbent or antibiotic-inactivating enzyme is administered at least once daily, in particular at least twice daily, more in particular three times daily or four times daily. In other particular embodiments, the adsorbent or antibiotic inactivating enzyme is administered throughout the treatment with the agent causing dysbiosis, in particular with an antibiotic. In a variant of this embodiment, the adsorbent or antibiotic-inactivating enzyme may be administered for a longer period of time than the dysbiosis-causing agent, in particular than the antibiotic, in order to ensure elimination of any residual dysbiosis-causing agent, in particular any residual antibiotic. For example, the adsorbent or antibiotic-inactivating enzyme may still be administered at least one day, e.g. two days, after discontinuing the administration of the agent causing dysbiosis, in particular the administration of the antibiotic.

In a particular embodiment, the invention relates to an adsorbent or antibiotic inactivating enzyme which is used in combination with, in particular almost simultaneously with, an antibiotic for a subject in need of transplantation. According to this embodiment, the adsorbent or antibiotic-inactivating enzyme prevents the potentially adverse effect of the antibiotic on the subject's intestinal microbial flora and thus can treat or prevent GVHD.

In particular embodiments, the invention may be suitably used in patients at risk of GVHD, for example patients taking antibiotics waiting for an HSCT procedure, to prevent GVHD from occurring or to reduce the severity of GVHD episodes if one episode occurs despite the start of the treatment of the invention.

In particular, the invention is useful for patients awaiting or during an HSCT procedure when receiving antibiotics, particularly in the neutropenic stage. The invention can also be used in these patients when they receive antibiotics prior to the neutropenic stage in order to maintain optimal microbial flora balance. The invention can also be used for patients diagnosed with cancer of the blood or bone marrow when they receive antibiotics in order to maintain the microbial flora for the longest time possible in the best possible state and to improve the outcome of HSCT in case a HSCT procedure is deemed necessary for curing said patient.

The invention can also be used for patients who have received an HSCT procedure when they receive antibiotics in order to prevent the occurrence of GVHD syndrome or to avoid worsening of the disease if the patient already suffers from acute or chronic GVHD.

In particular embodiments, the present invention may be used each time the subject takes an antibiotic. The invention may also be used after the subject has received fecal microbial transplantation or treatment with probiotics to restore its microbial flora diversity and is at risk for GVHD.

In particular embodiments, the subject is administered with an immunosuppressive agent such as methotrexate, tacrolimus, everolimus, sirolimus, mycophenolate mofetil, or cyclosporine a. In another specific embodiment, the subject is administered an anti-inflammatory drug, such as a corticosteroid.

In other particular embodiments, the subject has fever. In particular, the antibiotic to be eliminated from the intestinal tract of the subject is prescribed due to the fever.

In other particular embodiments, the active compounds of the invention are used in a method for preventing alteration of the microbial flora in a subject who has received, or will receive an allograft.

The invention may also be used in subjects with a high risk of GVHD, e.g. subjects with a previous GVHD episode in the years before a new antibiotic cure, a new hospitalization or a new immunosuppressive cure.

Thus, the invention also relates to a kit comprising an adsorbent and an agent causing dysbiosis, such as an antibiotic, or to a kit comprising an antibiotic inactivating enzyme and an antibiotic. The kit may be used to treat or prevent a pathological condition that may be treated or prevented using the dysbiosis-causing agent, e.g., an antibiotic. In a particular embodiment of the kit, the agent causing dysbiosis is an antibiotic. The kit may further comprise instructions for carrying out the method of the invention for treating or preventing GVHD. The components of the kit may be administered simultaneously, separately or sequentially. As provided above, the adsorbent or antibiotic inactivating enzyme may in particular be administered before, during or after administration of the agent causing dysbiosis, e.g. antibiotic, in particular shortly before or shortly after, more in particular shortly before.

Other aspects and advantages of the invention will be disclosed in the following illustrative experimental section.

Examples

Example 1:

to assess the clinical status of mice after HSCT and notably detect GVHD cases, a scoring system can be used to monitor multiple clinical symptoms. The scores take into account body weight, presence of diarrhea, dehydration, hair loss, and ability to move. For each criterion, a score is defined between 0, 1 or 2. The total score is obtained by adding the scores of each criterion. Healthy mice have a low total score, while unhealthy mice have a high total score and have a high score in one or more criteria.

If there is no weight loss or less than 10% weight loss, this criterion scores 0. If the weight loss was between 10 and 20%, the score was 1. If the weight loss is above 20%, a score of 2 is given.

If the mice did not exhibit diarrhea, they were scored as 0. If mild diarrhea is observed, a score of 1 is given. If diarrhea is severe, it is scored as 2.

For dehydration, a score of 0 is given if no signs of dehydration are observed, 1 if there is slight dehydration, and 2 if dehydration is severe.

If no depilation is present, a score of 0 is given. If there was mild alopecia, the score was 1. If the epilation was severe, it was scored as 2.

If the mice exhibited normal exploratory movement, the score was 0. If the movement of the mouse is limited, the score is 1. If the mice were barely moving, the score was 2.

Other criteria, such as clinical observations of the eye, ear, back, fur, or other clinically relevant results, may be added to the score, if desired.

Example 2:

to assess the effect of antibiotic use on the development of GvHD following HSCT, female BALB/C mice were irradiated with a gamma-ray source (8Gy) on day 0 and 5x10 from C57BI/6 mice on day +1⁶Spleen cells and 1x10⁷A mixture of individual Bone Marrow (BM) cells was injected intravenously. The mice were provided with antibiotics or placebo by subcutaneous administration from day-7 to day + 20. During the experiment, survival, body weight and clinical scores as defined in example 1 were recorded daily until day + 30. High clinical scores are associated with GVHD complications. On day +30, a higher disease score was observed in the mice receiving the antibiotic compared to the mice not receiving the antibiotic.

Example 3:

to evaluate the effect of the adsorbent administered with the antibiotic on the occurrence of GvHD after HSCT, the same procedure as in example 2 was used and for each group treated with the antibiotic, another group additionally receiving the adsorbent twice daily through an oral feeding tube from day-7 to day +25 was added. On day +30, a lower disease score was observed in mice receiving the adsorbent compared to mice receiving the antibiotic without the adsorbent.

Example 4:

to evaluate the effect of antibiotic inactivating enzymes administered with antibiotics on the occurrence of GvHD after HSCT, the same procedure as in example 2 was performed using β -lactam antibiotics. An additional group was introduced in which animals were also dosed twice daily with beta-lactamase provided via oral feeding tubes from day-7 to day + 25. On day +30, a lower disease score was observed in mice receiving the beta-lactamase compared to mice receiving the antibiotic without the beta-lactamase.

Example 5:

to investigate the effect of adsorbents on the prevention or attenuation of acute graft versus host disease, we performed the following experiments.

30 mice (129, female) were lethally irradiated on day 0 and 5 million C57 BL/6T-cell exfoliated bone marrow cells and 1 million C57BL/6 spleen T cells were transplanted.

The mice were then divided into 3 groups. In group a, they received no further treatment. In group B, imipenem treatment was started on day 10. It consists of 3 times a week administration of imipenem (100mg/kg) by subcutaneous route. Imipenem treatment was stopped on day 21. In group C, the same imipenem treatment as in group B was performed. In addition, group C mice received activated charcoal (14.4mg/g) mixed with their diet in hydrogel form daily from day 9 to day 22.

The clinical status of the mice was monitored daily from day 1 to day 21. In the context of such hematopoietic stem cell transplantation, mice are known to have a high propensity to develop lethal acute graft-versus-host disease.

The percent survival at day 21 in each group is presented in the table below:

group of	Survival at day 21
		A (bone marrow + T cell transplantation)	70％
B (bone marrow + T cell transplantation + Imipenem)	60％
		C (bone marrow + T cell transplantation + Imipenem + activated carbon)	90％

As presented in the present invention, it was observed that the addition of antibiotics to the mouse treatment regimen caused a potentiation of the GvHD condition and a higher mortality rate. Surprisingly, the addition of activated carbon, an adsorbent capable of capturing imipenem in the intestinal microflora of mice, reverses the deleterious effects of the antibiotic and can even improve the results of the mice by even further reducing the mortality caused by graft-versus-host disease.

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210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 9

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 Q34R E130D Q22N

<400> 9

Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 10

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 Q34R E130D Q22H

<400> 10

Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 11

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 Q34R Q22H DCT236

<400> 11

Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 12

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 Q34R Q22N DCT236

<400> 12

Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 13

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 Q34R E130D DCT236

<400> 13

Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 14

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 Q34R E130D Q22N DCT236

<400> 14

Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 15

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 Q34R E130D Q22H DCT236

<400> 15

Val Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 16

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V1M Q34R Q22H

<400> 16

Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 17

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V1M Q34R Q22N

<400> 17

Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 18

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V1M Q34R E130D

<400> 18

Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 19

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V1M Q34R E130D Q22N

<400> 19

Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 20

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V1M Q34R E130D Q22H

<400> 20

Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 21

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V1M Q34R Q22H DCT236

<400> 21

Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 22

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V1M Q34R Q22N DCT236

<400> 22

Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 23

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V1M Q34R E130D DCT236

<400> 23

Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 24

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V1M Q34R E130D Q22N DCT236

<400> 24

Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 25

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V1M Q34R E130D Q22H DCT236

<400> 25

Met Asp Ser Ser Gly Glu Tyr Pro Thr Val Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 26

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A

<400> 26

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Gln Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 27

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A Q34R Q22H

<400> 27

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 28

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A Q34R Q22N

<400> 28

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 29

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A Q34R E130D

<400> 29

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 30

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A Q34R E130D Q22N

<400> 30

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 31

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A Q34R E130D Q22H

<400> 31

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 32

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A Q34R Q22H DCT236

<400> 32

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 33

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A Q34R Q22N DCT236

<400> 33

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 34

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A Q34R E130D DCT236

<400> 34

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 35

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A Q34R E130D Q22N DCT236

<400> 35

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 36

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A Q34R E130D Q22H DCT236

<400> 36

Val Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 37

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A V1M Q34R Q22H

<400> 37

Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 38

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A V1M Q34R Q22N

<400> 38

Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 39

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A V1M Q34R E130D

<400> 39

Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 40

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A V1M Q34R E130D Q22N

<400> 40

Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 41

<211> 240

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A V1M Q34R E130D Q22H

<400> 41

Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn Arg Ser Val Val Glu

225 230 235 240

<210> 42

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A V1M Q34R Q22H DCT236

<400> 42

Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 43

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A V1M Q34R Q22N DCT236

<400> 43

Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Glu Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 44

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A V1M Q34R E130D DCT236

<400> 44

Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Gln Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 45

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A V1M Q34R E130D Q22N DCT236

<400> 45

Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr Asn Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 46

<211> 235

<212> PRT

<213> Artificial

<220>

<223> VIM-2 V10A V1M Q34R E130D Q22H DCT236

<400> 46

Met Asp Ser Ser Gly Glu Tyr Pro Thr Ala Ser Glu Ile Pro Val Gly

1 5 10 15

Glu Val Arg Leu Tyr His Ile Ala Asp Gly Val Trp Ser His Ile Ala

20 25 30

Thr Arg Ser Phe Asp Gly Ala Val Tyr Pro Ser Asn Gly Leu Ile Val

35 40 45

Arg Asp Gly Asp Glu Leu Leu Leu Ile Asp Thr Ala Trp Gly Ala Lys

50 55 60

Asn Thr Ala Ala Leu Leu Ala Glu Ile Glu Lys Gln Ile Gly Leu Pro

65 70 75 80

Val Thr Arg Ala Val Ser Thr His Phe His Asp Asp Arg Val Gly Gly

85 90 95

Val Asp Val Leu Arg Ala Ala Gly Val Ala Thr Tyr Ala Ser Pro Ser

100 105 110

Thr Arg Arg Leu Ala Glu Val Glu Gly Asn Glu Ile Pro Thr His Ser

115 120 125

Leu Asp Gly Leu Ser Ser Ser Gly Asp Ala Val Arg Phe Gly Pro Val

130 135 140

Glu Leu Phe Tyr Pro Gly Ala Ala His Ser Thr Asp Asn Leu Val Val

145 150 155 160

Tyr Val Pro Ser Ala Ser Val Leu Tyr Gly Gly Cys Ala Ile Tyr Glu

165 170 175

Leu Ser Arg Thr Ser Ala Gly Asn Val Ala Asp Ala Asp Leu Ala Glu

180 185 190

Trp Pro Thr Ser Ile Glu Arg Ile Gln Gln His Tyr Pro Glu Ala Gln

195 200 205

Phe Val Ile Pro Gly His Gly Leu Pro Gly Gly Leu Asp Leu Leu Lys

210 215 220

His Thr Thr Asn Val Val Lys Ala His Thr Asn

225 230 235

<210> 47

<211> 263

<212> PRT

<213> Artificial

<220>

<223> P1A

<400> 47

Glu Met Lys Asp Asp Phe Ala Lys Leu Glu Glu Gln Phe Asp Ala Lys

1 5 10 15

Leu Gly Ile Phe Ala Leu Asp Thr Gly Thr Asn Arg Thr Val Ala Tyr

20 25 30

Arg Pro Asp Glu Arg Phe Ala Phe Ala Ser Thr Ile Lys Ala Leu Thr

35 40 45

Val Gly Val Leu Leu Gln Gln Lys Ser Ile Glu Asp Leu Asn Gln Arg

50 55 60

Ile Thr Tyr Thr Arg Asp Asp Leu Val Asn Tyr Asn Pro Ile Thr Glu

65 70 75 80

Lys His Val Asp Thr Gly Met Thr Leu Lys Glu Leu Ala Asp Ala Ser

85 90 95

Leu Arg Tyr Ser Asp Asn Ala Ala Gln Asn Leu Ile Leu Lys Gln Ile

100 105 110

Gly Gly Pro Glu Ser Leu Lys Lys Glu Leu Arg Lys Ile Gly Asp Glu

115 120 125

Val Thr Asn Pro Glu Arg Phe Glu Pro Glu Leu Asn Glu Val Asn Pro

130 135 140

Gly Glu Thr Gln Asp Thr Ser Thr Ala Arg Ala Leu Val Thr Ser Leu

145 150 155 160

Arg Ala Phe Ala Leu Glu Asp Lys Leu Pro Ser Glu Lys Arg Glu Leu

165 170 175

Leu Ile Asp Trp Met Lys Arg Asn Thr Thr Gly Asp Ala Leu Ile Arg

180 185 190

Ala Gly Val Pro Asp Gly Trp Glu Val Ala Asp Lys Thr Gly Ala Ala

195 200 205

Ser Tyr Gly Thr Arg Asn Asp Ile Ala Ile Ile Trp Pro Pro Lys Gly

210 215 220

Asp Pro Val Val Leu Ala Val Leu Ser Ser Arg Asp Lys Lys Asp Ala

225 230 235 240

Lys Tyr Asp Asp Lys Leu Ile Ala Glu Ala Thr Lys Val Val Met Lys

245 250 255

Ala Leu Asn Met Asn Gly Lys

260

<210> 48

<211> 215

<212> PRT

<213> Artificial

<220>

<223> P2A

<400> 48

Glu Thr Gly Thr Ile Ser Ile Ser Gln Leu Asn Lys Asn Val Trp Val

1 5 10 15

His Thr Glu Leu Gly Tyr Phe Asn Gly Glu Ala Val Pro Ser Asn Gly

20 25 30

Leu Val Leu Asn Thr Ser Lys Gly Leu Val Leu Val Asp Ser Ser Trp

35 40 45

Asp Asn Lys Leu Thr Lys Glu Leu Ile Glu Met Val Glu Lys Lys Phe

50 55 60

Gln Lys Arg Val Thr Asp Val Ile Ile Thr His Ala His Ala Asp Arg

65 70 75 80

Ile Gly Gly Ile Thr Ala Leu Lys Glu Arg Gly Ile Lys Ala His Ser

85 90 95

Thr Ala Leu Thr Ala Glu Leu Ala Lys Asn Ser Gly Tyr Glu Glu Pro

100 105 110

Leu Gly Asp Leu Gln Thr Ile Thr Ser Leu Lys Phe Gly Asn Thr Lys

115 120 125

Val Glu Thr Phe Tyr Pro Gly Lys Gly His Thr Glu Asp Asn Ile Val

130 135 140

Val Trp Leu Pro Gln Tyr Gln Ile Leu Ala Gly Gly Cys Leu Val Lys

145 150 155 160

Ser Ala Glu Ala Lys Asp Leu Gly Asn Val Ala Asp Ala Tyr Val Asn

165 170 175

Glu Trp Ser Thr Ser Ile Glu Asn Val Leu Lys Arg Tyr Gly Asn Ile

180 185 190

Asn Ser Trp Pro Gly His Gly Glu Val Gly Asp Lys Gly Leu Leu Leu

195 200 205

His Thr Leu Asp Leu Leu Lys

210 215

<210> 49

<211> 258

<212> PRT

<213> Artificial

<220>

<223> P3A variants

<220>

<221> MISC_FEATURE

<222> (243)..(243)

<223> Xaa can be any naturally occurring amino acid

<400> 49

Met Lys Asp Asp Phe Ala Lys Leu Glu Glu Gln Phe Asp Ala Lys Leu

1 5 10 15

Gly Ile Phe Ala Leu Asp Thr Gly Thr Asn Arg Thr Val Ala Tyr Arg

20 25 30

Pro Asp Glu Arg Phe Ala Phe Ala Ser Thr Ile Lys Ala Leu Thr Val

35 40 45

Gly Val Leu Leu Gln Gln Lys Ser Ile Glu Asp Leu Asn Gln Arg Ile

50 55 60

Thr Tyr Thr Arg Asp Asp Leu Val Asn Tyr Asn Pro Ile Thr Glu Lys

65 70 75 80

His Val Asp Thr Gly Met Thr Leu Lys Glu Leu Ala Asp Ala Ser Leu

85 90 95

Arg Tyr Ser Asp Asn Ala Ala Gln Asn Leu Ile Leu Lys Gln Ile Gly

100 105 110

Gly Pro Glu Ser Leu Lys Lys Glu Leu Arg Lys Ile Gly Asp Glu Val

115 120 125

Thr Asn Pro Glu Arg Phe Glu Pro Glu Leu Asn Glu Val Asn Pro Gly

130 135 140

Glu Thr Gln Asp Thr Ser Thr Ala Arg Ala Leu Val Thr Ser Leu Arg

145 150 155 160

Ala Phe Ala Leu Glu Asp Lys Leu Pro Ser Glu Lys Arg Glu Leu Leu

165 170 175

Ile Asp Trp Met Lys Arg Asn Thr Thr Gly Asp Ala Leu Ile Arg Ala

180 185 190

Gly Val Pro Asp Gly Trp Glu Val Ala Asp Lys Thr Gly Ala Ala Ser

195 200 205

Tyr Gly Thr Arg Asn Asp Ile Ala Ile Ile Trp Pro Pro Lys Gly Asp

210 215 220

Pro Val Val Leu Ala Val Leu Ser Ser Arg Asp Lys Lys Asp Ala Lys

225 230 235 240

Tyr Asp Xaa Lys Leu Ile Ala Glu Ala Thr Lys Val Val Met Lys Ala

245 250 255

Leu Asn

<210> 50

<211> 299

<212> PRT

<213> Artificial

<220>

<223> P3A variants

<220>

<221> misc_feature

<222> (280)..(280)

<223> Xaa can be any naturally occurring amino acid

<220>

<221> MISC_FEATURE

<222> (290)..(290)

<223> Xaa can be any naturally occurring amino acid

<400> 50

Met Ile Gln Lys Arg Lys Arg Thr Val Ser Phe Arg Leu Val Leu Met

1 5 10 15

Cys Thr Leu Leu Phe Val Ser Leu Pro Ile Thr Lys Thr Ser Ala Gln

20 25 30

Ala Ser Lys Thr Glu Met Lys Asp Asp Phe Ala Lys Leu Glu Glu Gln

35 40 45

Phe Asp Ala Lys Leu Gly Ile Phe Ala Leu Asp Thr Gly Thr Asn Arg

50 55 60

Thr Val Ala Tyr Arg Pro Asp Glu Arg Phe Ala Phe Ala Ser Thr Ile

65 70 75 80

Lys Ala Leu Thr Val Gly Val Leu Leu Gln Gln Lys Ser Ile Glu Asp

85 90 95

Leu Asn Gln Arg Ile Thr Tyr Thr Arg Asp Asp Leu Val Asn Tyr Asn

100 105 110

Pro Ile Thr Glu Lys His Val Asp Thr Gly Met Thr Leu Lys Glu Leu

115 120 125

Ala Asp Ala Ser Leu Arg Tyr Ser Asp Asn Ala Ala Gln Asn Leu Ile

130 135 140

Leu Lys Gln Ile Gly Gly Pro Glu Ser Leu Lys Lys Glu Leu Arg Lys

145 150 155 160

Ile Gly Asp Glu Val Thr Asn Pro Glu Arg Phe Glu Pro Glu Leu Asn

165 170 175

Glu Val Asn Pro Gly Glu Thr Gln Asp Thr Ser Thr Ala Arg Ala Leu

180 185 190

Val Thr Ser Leu Arg Ala Phe Ala Leu Glu Asp Lys Leu Pro Ser Glu

195 200 205

Lys Arg Glu Leu Leu Ile Asp Trp Met Lys Arg Asn Thr Thr Gly Asp

210 215 220

Ala Leu Ile Arg Ala Gly Val Pro Asp Gly Trp Glu Val Ala Asp Lys

225 230 235 240

Thr Gly Ala Ala Ser Tyr Gly Thr Arg Asn Asp Ile Ala Ile Ile Trp

245 250 255

Pro Pro Lys Gly Asp Pro Val Val Leu Ala Val Leu Ser Ser Arg Asp

260 265 270

Lys Lys Asp Ala Lys Tyr Asp Xaa Lys Leu Ile Ala Glu Ala Thr Lys

275 280 285

Val Val Met Lys Ala Leu Asn Met Asn Gly Lys

290 295

<210> 51

<211> 293

<212> PRT

<213> Artificial

<220>

<223> KPC-1/2

<400> 51

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Ser Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro

245 250 255

Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His

260 265 270

Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu

275 280 285

Gly Val Asn Gly Gln

290

<210> 52

<211> 293

<212> PRT

<213> Artificial

<220>

<223> KPC-3

<400> 52

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro

245 250 255

Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr

260 265 270

Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu

275 280 285

Gly Val Asn Gly Gln

290

<210> 53

<211> 292

<212> PRT

<213> Artificial

<220>

<223> KPC-4

<400> 53

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Arg Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Gly Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Trp Trp Pro Thr Gly Arg Ala Pro Ile

245 250 255

Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His Ser

260 265 270

Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu Gly

275 280 285

Val Asn Gly Gln

290

<210> 54

<211> 293

<212> PRT

<213> Artificial

<220>

<223> KPC-5

<400> 54

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Arg Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro

245 250 255

Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His

260 265 270

Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu

275 280 285

Gly Val Asn Gly Gln

290

<210> 55

<211> 292

<212> PRT

<213> Artificial

<220>

<223> KPC-6

<400> 55

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Gly Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Trp Trp Pro Thr Gly Arg Ala Pro Ile

245 250 255

Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His Ser

260 265 270

Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu Gly

275 280 285

Val Asn Gly Gln

290

<210> 56

<211> 293

<212> PRT

<213> Artificial

<220>

<223> KPC-7

<400> 56

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Ile Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Leu Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro

245 250 255

Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr

260 265 270

Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu

275 280 285

Gly Val Asn Gly Gln

290

<210> 57

<211> 292

<212> PRT

<213> Artificial

<220>

<223> KPC-8

<400> 57

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Gly Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Trp Trp Pro Thr Gly Arg Ala Pro Ile

245 250 255

Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr Ser

260 265 270

Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu Gly

275 280 285

Val Asn Gly Gln

290

<210> 58

<211> 293

<212> PRT

<213> Artificial

<220>

<223> KPC-9

<400> 58

Met Ser Lys Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Ala Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro

245 250 255

Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr

260 265 270

Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu

275 280 285

Gly Val Asn Gly Gln

290

<210> 59

<211> 293

<212> PRT

<213> Artificial

<220>

<223> KPC-10

<400> 59

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Arg Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro

245 250 255

Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr

260 265 270

Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu

275 280 285

Gly Val Asn Gly Gln

290

<210> 60

<211> 293

<212> PRT

<213> Artificial

<220>

<223> KPC-11

<400> 60

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Leu Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro

245 250 255

Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His

260 265 270

Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu

275 280 285

Gly Val Asn Gly Gln

290

<210> 61

<211> 293

<212> PRT

<213> Artificial

<220>

<223> KPC-12

<400> 61

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Met Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro

245 250 255

Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His

260 265 270

Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu

275 280 285

Gly Val Asn Gly Gln

290

<210> 62

<211> 293

<212> PRT

<213> Artificial

<220>

<223> KPC-13

<400> 62

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Gly Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro

245 250 255

Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr

260 265 270

Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu

275 280 285

Gly Val Asn Gly Gln

290

<210> 63

<211> 291

<212> PRT

<213> Artificial

<220>

<223> KPC-14

<400> 63

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr

225 230 235 240

Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro Ile Val

245 250 255

Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His Ser Glu

260 265 270

Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu Gly Val

275 280 285

Asn Gly Gln

290

<210> 64

<211> 292

<212> PRT

<213> Artificial

<220>

<223> KPC-15

<400> 64

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Arg Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Leu Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Lys Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Phe Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Gly Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Trp Trp Pro Thr Gly Arg Ala Pro Ile

245 250 255

Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys Tyr Ser

260 265 270

Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu Gly

275 280 285

Val Asn Gly Gln

290

<210> 65

<211> 293

<212> PRT

<213> Artificial

<220>

<223> KPC-17

<400> 65

Met Ser Leu Tyr Arg Arg Leu Val Leu Leu Ser Cys Leu Ser Trp Pro

1 5 10 15

Leu Ala Gly Phe Ser Ala Thr Ala Leu Thr Asn Leu Val Ala Glu Pro

20 25 30

Phe Ala Lys Leu Glu Gln Asp Phe Gly Gly Ser Ile Gly Val Tyr Ala

35 40 45

Met Asp Thr Gly Ser Gly Ala Thr Val Ser Tyr Arg Ala Glu Glu Arg

50 55 60

Phe Pro Leu Cys Ser Ser Phe Lys Gly Phe Leu Ala Ala Ala Val Leu

65 70 75 80

Ala Arg Ser Gln Gln Gln Ala Gly Leu Leu Asp Thr Pro Ile Arg Tyr

85 90 95

Gly Lys Asn Ala Leu Val Pro Trp Ser Pro Ile Ser Glu Lys Tyr Leu

100 105 110

Thr Thr Gly Met Thr Val Ala Glu Leu Ser Ala Ala Ala Val Gln Tyr

115 120 125

Ser Asp Asn Ala Ala Ala Asn Leu Leu Leu Lys Glu Leu Gly Gly Pro

130 135 140

Ala Gly Leu Thr Ala Phe Met Arg Ser Ile Gly Asp Thr Thr Phe Arg

145 150 155 160

Leu Asp Arg Trp Glu Leu Glu Leu Asn Ser Ala Ile Pro Gly Asp Ala

165 170 175

Arg Asp Thr Ser Ser Pro Arg Ala Val Thr Glu Ser Leu Gln Lys Leu

180 185 190

Thr Leu Gly Ser Ala Leu Ala Ala Pro Gln Arg Gln Gln Leu Val Asp

195 200 205

Trp Leu Lys Gly Asn Thr Thr Gly Asn His Arg Ile Arg Ala Ala Val

210 215 220

Pro Ala Asp Trp Ala Val Gly Asp Lys Thr Gly Thr Cys Gly Val Tyr

225 230 235 240

Gly Thr Ala Asn Asp Tyr Ala Val Val Trp Pro Thr Gly Arg Ala Pro

245 250 255

Ile Val Leu Ala Val Tyr Thr Arg Ala Pro Asn Lys Asp Asp Lys His

260 265 270

Ser Glu Ala Val Ile Ala Ala Ala Ala Arg Leu Ala Leu Glu Gly Leu

275 280 285

Gly Val Asn Gly Gln

290

<210> 66

<211> 269

<212> PRT

<213> Artificial

<220>

<223> NDM-1

<400> 66

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Met Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn Phe

165 170 175

Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn

180 185 190

Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu

195 200 205

Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp

210 215 220

Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro

225 230 235 240

Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala

245 250 255

Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 67

<211> 269

<212> PRT

<213> Artificial

<220>

<223> NDM-2

<400> 67

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Ala Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Met Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn Phe

165 170 175

Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn

180 185 190

Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu

195 200 205

Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp

210 215 220

Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro

225 230 235 240

Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala

245 250 255

Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 68

<211> 269

<212> PRT

<213> Artificial

<220>

<223> NDM-3

<400> 68

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asn Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Met Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn Phe

165 170 175

Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn

180 185 190

Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu

195 200 205

Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp

210 215 220

Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro

225 230 235 240

Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala

245 250 255

Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 69

<211> 268

<212> PRT

<213> Artificial

<220>

<223> NDM-4

<400> 69

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly

165 170 175

Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile

180 185 190

Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile

195 200 205

Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp Thr

210 215 220

Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys

225 230 235 240

Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala

245 250 255

Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 70

<211> 270

<212> PRT

<213> Artificial

<220>

<223> NDM-5

<400> 70

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Leu Val Asp Thr Ala Trp Thr Asp Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Val Val Thr His Ala His Gln Asp Lys Met Gly Gly

115 120 125

Met Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu

130 135 140

Ser Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser

145 150 155 160

Leu Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn

165 170 175

Phe Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp

180 185 190

Asn Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys

195 200 205

Leu Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala

210 215 220

Asp Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe

225 230 235 240

Pro Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg

245 250 255

Ala Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265 270

<210> 71

<211> 269

<212> PRT

<213> Artificial

<220>

<223> NDM-6

<400> 71

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Met Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn Phe

165 170 175

Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn

180 185 190

Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu

195 200 205

Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp

210 215 220

Thr Glu His Tyr Ala Ala Ser Val Arg Ala Phe Gly Ala Ala Phe Pro

225 230 235 240

Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala

245 250 255

Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 72

<211> 268

<212> PRT

<213> Artificial

<220>

<223> NDM-7

<400> 72

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asn Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly

165 170 175

Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile

180 185 190

Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile

195 200 205

Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp Thr

210 215 220

Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys

225 230 235 240

Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala

245 250 255

Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 73

<211> 269

<212> PRT

<213> Artificial

<220>

<223> NDM-8

<400> 73

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Gly Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Val Trp Glu Pro Ala Thr Ala Pro Asn Phe

165 170 175

Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn

180 185 190

Ile Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu

195 200 205

Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp

210 215 220

Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro

225 230 235 240

Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala

245 250 255

Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 74

<211> 268

<212> PRT

<213> Artificial

<220>

<223> NDM-9

<400> 74

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Lys Gly Met Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly

165 170 175

Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile

180 185 190

Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile

195 200 205

Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp Thr

210 215 220

Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys

225 230 235 240

Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala

245 250 255

Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 75

<211> 269

<212> PRT

<213> Artificial

<220>

<223> NDM-10

<400> 75

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Ser

20 25 30

Pro Thr Ile Asp Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Ser Phe Gly Ala Val Thr Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Trp Asp Thr Ala Trp Thr Asp Asp Gln

85 90 95

Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro Val

100 105 110

Ala Leu Ala Val Val Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Met Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Trp Val Glu Pro Ala Thr Ala Pro Asn Phe

165 170 175

Gly Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn

180 185 190

Ile Thr Val Gly Ile Asp Arg Thr Asp Ile Ala Phe Gly Gly Cys Leu

195 200 205

Ile Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp

210 215 220

Thr Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro

225 230 235 240

Lys Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala

245 250 255

Ala Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 76

<211> 268

<212> PRT

<213> Artificial

<220>

<223> NDM-11

<400> 76

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly

165 170 175

Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile

180 185 190

Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile

195 200 205

Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Asp Asp Ala Asp Thr

210 215 220

Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys

225 230 235 240

Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala

245 250 255

Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 77

<211> 268

<212> PRT

<213> Artificial

<220>

<223> NDM-12

<400> 77

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asp Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly

165 170 175

Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile

180 185 190

Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile

195 200 205

Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Asp Asp Ala Asp Thr

210 215 220

Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys

225 230 235 240

Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala

245 250 255

Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 78

<211> 268

<212> PRT

<213> Artificial

<220>

<223> NDM-13

<400> 78

Met Glu Leu Pro Asn Ile Met His Pro Val Ala Lys Leu Ser Thr Ala

1 5 10 15

Leu Ala Ala Ala Leu Met Leu Ser Gly Cys Met Pro Gly Glu Ile Arg

20 25 30

Pro Thr Ile Gly Gln Gln Met Glu Thr Gly Asp Gln Arg Phe Gly Asp

35 40 45

Leu Val Phe Arg Gln Leu Ala Pro Asn Val Trp Gln His Thr Ser Tyr

50 55 60

Leu Asp Met Pro Gly Phe Gly Ala Val Ala Ser Asn Gly Leu Ile Val

65 70 75 80

Arg Asp Gly Gly Arg Val Leu Val Val Asp Thr Ala Trp Thr Asn Asp

85 90 95

Gln Thr Ala Gln Ile Leu Asn Trp Ile Lys Gln Glu Ile Asn Leu Pro

100 105 110

Val Ala Leu Ala Trp Thr His Ala His Gln Asp Lys Met Gly Gly Met

115 120 125

Asp Ala Leu His Ala Ala Gly Ile Ala Thr Tyr Ala Asn Ala Leu Ser

130 135 140

Asn Gln Leu Ala Pro Gln Glu Gly Leu Val Ala Ala Gln His Ser Leu

145 150 155 160

Thr Phe Ala Ala Asn Gly Trp Glu Pro Ala Thr Ala Pro Asn Phe Gly

165 170 175

Pro Leu Lys Val Phe Tyr Pro Gly Pro Gly His Thr Ser Asp Asn Ile

180 185 190

Thr Val Gly Ile Asp Gly Thr Asp Ile Ala Phe Gly Gly Cys Leu Ile

195 200 205

Lys Asp Ser Lys Ala Lys Ser Leu Gly Asn Leu Gly Asp Ala Asp Thr

210 215 220

Glu His Tyr Ala Ala Ser Ala Arg Ala Phe Gly Ala Ala Phe Pro Lys

225 230 235 240

Ala Ser Met Ile Val Met Ser His Ser Ala Pro Asp Ser Arg Ala Ala

245 250 255

Ile Thr His Thr Ala Arg Met Ala Asp Lys Leu Arg

260 265

<210> 79

<211> 260

<212> PRT

<213> Artificial

<220>

<223> P4A

<400> 79

Glu Met Lys Asp Asp Phe Ala Lys Leu Glu Glu Gln Phe Asp Ala Lys

1 5 10 15

Leu Gly Ile Phe Ala Leu Asp Thr Gly Thr Asn Arg Thr Val Ala Tyr

20 25 30

Arg Pro Asp Glu Arg Phe Ala Phe Ala Ser Thr Ile Lys Ala Leu Thr

35 40 45

Val Gly Val Leu Leu Gln Gln Lys Ser Ile Glu Asp Leu Asn Gln Arg

50 55 60

Ile Thr Thr Arg Asp Asp Leu Val Asn Tyr Asn Pro Ile Thr Glu Lys

65 70 75 80

His Val Asp Thr Gly Met Thr Leu Lys Glu Leu Ala Asp Ala Ser Leu

85 90 95

Arg Tyr Ser Asp Asn Ala Ala Gln Asn Leu Ile Leu Lys Gln Ile Gly

100 105 110

Gly Pro Glu Ser Leu Lys Lys Glu Leu Arg Lys Ile Gly Asp Glu Val

115 120 125

Thr Asn Pro Glu Arg Phe Glu Pro Glu Leu Asn Glu Val Asn Pro Gly

130 135 140

Glu Thr Gln Asp Thr Ser Thr Ala Arg Ala Leu Val Thr Ser Leu Arg

145 150 155 160

Ala Phe Ala Leu Glu Asp Lys Leu Pro Ser Glu Lys Arg Glu Leu Leu

165 170 175

Ile Asp Trp Met Lys Arg Asn Thr Thr Gly Asp Ala Leu Ile Arg Ala

180 185 190

Gly Val Pro Asp Gly Trp Glu Val Gly Asp Lys Thr Gly Ser Gly Asp

195 200 205

Tyr Gly Thr Arg Asn Asp Ile Ala Ile Ile Trp Pro Pro Lys Gly Asp

210 215 220

Pro Trp Leu Ala Val Leu Ser Ser Arg Asp Lys Lys Asp Ala Lys Tyr

225 230 235 240

Asp Asn Lys Leu Ile Ala Glu Ala Thr Lys Trp Met Lys Ala Leu Asn

245 250 255

Met Asn Gly Lys

260

<210> 80

<211> 299

<212> PRT

<213> Artificial

<220>

<223> P4A + signal peptide

<400> 80

Met Ile Gln Lys Arg Lys Arg Thr Val Ser Phe Arg Leu Val Leu Met

1 5 10 15

Cys Thr Leu Leu Phe Val Ser Leu Pro Ile Thr Lys Thr Ser Ala Gln

20 25 30

Ala Ser Lys Thr Glu Met Lys Asp Asp Phe Ala Lys Leu Glu Glu Gln

35 40 45

Phe Asp Ala Lys Leu Gly Ile Phe Ala Leu Asp Thr Gly Thr Asn Arg

50 55 60

Thr Val Ala Tyr Arg Pro Asp Glu Arg Phe Ala Phe Ala Ser Thr Ile

65 70 75 80

Lys Ala Leu Thr Val Gly Val Leu Leu Gln Gln Lys Ser Ile Glu Asp

85 90 95

Leu Asn Gln Arg Ile Thr Tyr Thr Arg Asp Asp Leu Val Asn Tyr Asn

100 105 110

Pro Ile Thr Glu Lys His Val Asp Thr Gly Met Thr Leu Lys Glu Leu

115 120 125

Ala Asp Ala Ser Leu Arg Tyr Ser Asp Asn Ala Ala Gln Asn Leu Ile

130 135 140

Leu Lys Gln Ile Gly Gly Pro Glu Ser Leu Lys Lys Glu Leu Arg Lys

145 150 155 160

Ile Gly Asp Glu Val Thr Asn Pro Glu Arg Phe Glu Pro Glu Leu Asn

165 170 175

Glu Val Asn Pro Gly Glu Thr Gln Asp Thr Ser Thr Ala Arg Ala Leu

180 185 190

Val Thr Ser Leu Arg Ala Phe Ala Leu Glu Asp Lys Leu Pro Ser Glu

195 200 205

Lys Arg Glu Leu Leu Ile Asp Trp Met Lys Arg Asn Thr Thr Gly Asp

210 215 220

Ala Leu Ile Arg Ala Gly Val Pro Asp Gly Trp Glu Val Gly Asp Lys

225 230 235 240

Thr Gly Ser Gly Asp Tyr Gly Thr Arg Asn Asp Ile Ala Ile Ile Trp

245 250 255

Pro Pro Lys Gly Asp Pro Val Val Leu Ala Val Leu Ser Ser Arg Asp

260 265 270

Lys Lys Asp Ala Lys Tyr Asp Asn Lys Leu Ile Ala Glu Ala Thr Lys

275 280 285

Val Val Met Lys Ala Leu Asn Met Asn Gly Lys

290 295

<210> 81

<211> 263

<212> PRT

<213> Escherichia coli (Escherichia coli)

<400> 81

His Pro Glu Thr Leu Val Lys Val Lys Asp Ala Glu Asp Gln Leu Gly

1 5 10 15

Ala Arg Val Gly Tyr Ile Glu Leu Asp Leu Asn Ser Gly Lys Ile Leu

20 25 30

Glu Ser Phe Arg Pro Glu Glu Arg Phe Pro Met Val Ser Thr Phe Lys

35 40 45

Val Leu Leu Cys Gly Ala Val Leu Ser Arg Val Asp Ala Gly Gln Glu

50 55 60

Gln Leu Gly Arg Arg Ile His Tyr Ser Gln Asn Asp Leu Val Glu Tyr

65 70 75 80

Ser Pro Val Thr Glu Lys His Leu Thr Asp Gly Met Thr Val Arg Glu

85 90 95

Leu Cys Ser Ala Ala Ile Thr Met Ser Asp Asn Thr Ala Ala Asn Leu

100 105 110

Leu Leu Thr Thr Ile Gly Gly Pro Lys Glu Leu Thr Ala Phe Leu His

115 120 125

Asn Met Gly Asp His Val Thr Arg Leu Asp Arg Trp Glu Pro Glu Leu

130 135 140

Asn Glu Ala Ile Pro Asn Asp Glu Arg Asp Thr Thr Met Pro Ala Ala

145 150 155 160

Met Ala Thr Thr Leu Arg Lys Leu Leu Thr Gly Glu Leu Leu Thr Leu

165 170 175

Ala Ser Arg Gln Gln Leu Ile Asp Trp Met Glu Ala Asp Lys Val Ala

180 185 190

Gly Pro Leu Leu Arg Ser Ala Leu Pro Ala Gly Trp Phe Ile Ala Asp

195 200 205

Lys Ser Gly Ala Gly Glu Arg Gly Ser Arg Gly Ile Ile Ala Ala Leu

210 215 220

Gly Pro Asp Gly Lys Pro Ser Arg Ile Val Val Ile Tyr Thr Thr Gly

225 230 235 240

Ser Gln Ala Thr Met Asp Glu Arg Asp Arg Gln Ile Ala Glu Ile Gly

245 250 255

Ala Ser Leu Ile Lys His Trp

260

<210> 82

<211> 263

<212> PRT

<213> Escherichia coli (Escherichia coli)

<400> 82

Gln Thr Ser Ala Val Gln Gln Lys Leu Ala Ala Leu Glu Lys Ser Ser

1 5 10 15

Gly Gly Arg Leu Gly Val Ala Leu Ile Asp Thr Ala Asp Asn Thr Gln

20 25 30

Val Leu Tyr Arg Gly Asp Glu Arg Phe Pro Met Cys Ser Thr Ser Lys

35 40 45

Val Met Ala Ala Ala Ala Val Leu Lys Gln Ser Glu Thr Gln Lys Gln

50 55 60

Leu Leu Asn Gln Pro Val Glu Ile Lys Pro Ala Asp Leu Val Asn Tyr

65 70 75 80

Asn Pro Ile Ala Glu Lys His Val Asn Gly Thr Met Thr Leu Ala Glu

85 90 95

Leu Ser Ala Ala Ala Leu Gln Tyr Ser Asp Asn Thr Ala Met Asn Lys

100 105 110

Leu Ile Ala Gln Leu Gly Gly Pro Gly Gly Val Thr Ala Phe Ala Arg

115 120 125

Ala Ile Gly Asp Glu Thr Phe Arg Leu Asp Arg Thr Glu Pro Thr Leu

130 135 140

Asn Thr Ala Ile Pro Gly Asp Pro Arg Asp Thr Thr Thr Pro Arg Ala

145 150 155 160

Met Ala Gln Thr Leu Arg Gln Leu Thr Leu Gly His Ala Leu Gly Glu

165 170 175

Thr Gln Arg Ala Gln Leu Val Thr Trp Leu Lys Gly Asn Thr Thr Gly

180 185 190

Ala Ala Ser Ile Arg Ala Gly Leu Pro Thr Ser Trp Thr Ala Gly Asp

195 200 205

Lys Thr Gly Ser Gly Gly Tyr Gly Thr Thr Asn Asp Ile Ala Val Ile

210 215 220

Trp Pro Gln Gly Arg Ala Pro Leu Val Leu Val Thr Tyr Phe Thr Gln

225 230 235 240

Pro Gln Gln Asn Ala Glu Ser Arg Arg Asp Val Leu Ala Ser Ala Ala

245 250 255

Arg Ile Ile Ala Glu Gly Leu

260

<210> 83

<211> 257

<212> PRT

<213> Staphylococcus aureus (Staphylococcus aureus)

<400> 83

Lys Glu Leu Asn Asp Leu Glu Lys Lys Tyr Asn Ala His Ile Gly Val

1 5 10 15

Tyr Ala Leu Asp Thr Lys Ser Gly Lys Glu Val Lys Phe Asn Ser Asp

20 25 30

Lys Arg Phe Ala Tyr Ala Ser Thr Ser Lys Ala Ile Asn Ser Ala Ile

35 40 45

Leu Leu Glu Gln Val Pro Tyr Asn Lys Leu Asn Lys Lys Val His Ile

50 55 60

Asn Lys Asp Asp Ile Val Ala Tyr Ser Pro Ile Leu Glu Lys Tyr Val

65 70 75 80

Gly Lys Asp Ile Thr Leu Lys Ala Leu Ile Glu Ala Ser Met Thr Tyr

85 90 95

Ser Asp Asn Thr Ala Asn Asn Lys Ile Ile Lys Glu Ile Gly Gly Ile

100 105 110

Lys Lys Val Lys Gln Arg Leu Lys Glu Leu Gly Asp Lys Val Thr Asn

115 120 125

Pro Val Arg Tyr Glu Ile Glu Leu Asn Tyr Tyr Ser Pro Lys Ser Lys

130 135 140

Lys Asp Thr Ser Thr Pro Ala Ala Phe Gly Lys Thr Leu Asn Lys Leu

145 150 155 160

Ile Ala Asn Gly Lys Leu Ser Lys Glu Asn Lys Lys Phe Leu Leu Asp

165 170 175

Leu Met Leu Asn Asn Lys Ser Gly Asp Thr Leu Ile Lys Asp Gly Val

180 185 190

Pro Lys Asp Tyr Lys Val Ala Asp Lys Ser Gly Gln Ala Ile Thr Tyr

195 200 205

Ala Ser Arg Asn Asp Val Ala Phe Val Tyr Pro Lys Gly Gln Ser Glu

210 215 220

Pro Ile Val Leu Val Ile Phe Thr Asn Lys Asp Asn Lys Ser Asp Lys

225 230 235 240

Pro Asn Asp Lys Leu Ile Ser Glu Thr Ala Lys Ser Val Met Lys Glu

245 250 255

Phe

<210> 84

<211> 199

<212> PRT

<213> Escherichia coli (Escherichia coli)

<400> 84

Met Ser Asn Ala Lys Thr Lys Leu Gly Ile Thr Lys Tyr Ser Ile Val

1 5 10 15

Thr Asn Ser Asn Asp Ser Val Thr Leu Arg Leu Met Thr Glu His Asp

20 25 30

Leu Ala Met Leu Tyr Glu Trp Leu Asn Arg Ser His Ile Val Glu Trp

35 40 45

Trp Gly Gly Glu Glu Ala Arg Pro Thr Leu Ala Asp Val Gln Glu Gln

50 55 60

Tyr Leu Pro Ser Val Leu Ala Gln Glu Ser Val Thr Pro Tyr Ile Ala

65 70 75 80

Met Leu Asn Gly Glu Pro Ile Gly Tyr Ala Gln Ser Tyr Val Ala Leu

85 90 95

Gly Ser Gly Asp Gly Arg Trp Glu Glu Glu Thr Asp Pro Gly Val Arg

100 105 110

Gly Ile Asp Gln Leu Leu Ala Asn Ala Ser Gln Leu Gly Lys Gly Leu

115 120 125

Gly Thr Lys Leu Val Arg Ala Leu Val Glu Leu Leu Phe Asn Asp Pro

130 135 140

Glu Val Thr Lys Ile Gln Thr Asp Pro Ser Pro Ser Asn Leu Arg Ala

145 150 155 160

Ile Arg Cys Tyr Glu Lys Ala Gly Phe Glu Arg Gln Gly Thr Val Thr

165 170 175

Thr Pro Tyr Gly Pro Ala Val Tyr Met Val Gln Thr Arg Gln Ala Phe

180 185 190

Glu Arg Thr Arg Ser Asp Ala

195

<210> 85

<211> 419

<212> PRT

<213> Escherichia coli (Escherichia coli)

<400> 85

Met Arg Phe Glu Glu Trp Val Lys Asp Lys His Ile Pro Phe Lys Leu

1 5 10 15

Asn His Pro Asp Asp Asn Tyr Asp Asp Phe Lys Pro Leu Arg Lys Ile

20 25 30

Ile Gly Asp Thr Arg Val Val Ala Leu Gly Glu Asn Ser His Phe Ile

35 40 45

Lys Glu Phe Phe Leu Leu Arg His Thr Leu Leu Arg Phe Phe Ile Glu

50 55 60

Asp Leu Gly Phe Thr Thr Phe Ala Phe Glu Phe Gly Phe Ala Glu Gly

65 70 75 80

Gln Ile Ile Asn Asn Trp Ile His Gly Gln Gly Thr Asp Asp Glu Ile

85 90 95

Gly Arg Phe Leu Lys His Phe Tyr Tyr Pro Glu Glu Leu Lys Thr Thr

100 105 110

Phe Leu Trp Leu Arg Glu Tyr Asn Lys Ala Ala Lys Glu Lys Ile Thr

115 120 125

Phe Leu Gly Ile Asp Ile Pro Arg Asn Gly Gly Ser Tyr Leu Pro Asn

130 135 140

Met Glu Ile Val His Asp Phe Phe Arg Thr Ala Asp Lys Glu Ala Leu

145 150 155 160

His Ile Ile Asp Asp Ala Phe Asn Ile Ala Lys Lys Ile Asp Tyr Phe

165 170 175

Ser Thr Ser Gln Ala Ala Leu Asn Leu His Glu Leu Thr Asp Ser Glu

180 185 190

Lys Cys Arg Leu Thr Ser Gln Leu Ala Arg Val Lys Val Arg Leu Glu

195 200 205

Ala Met Ala Pro Ile His Ile Glu Lys Tyr Gly Ile Asp Lys Tyr Glu

210 215 220

Thr Ile Leu His Tyr Ala Asn Gly Met Ile Tyr Leu Asp Tyr Asn Ile

225 230 235 240

Gln Ala Met Ser Gly Phe Ile Ser Gly Gly Gly Met Gln Gly Asp Met

245 250 255

Gly Ala Lys Asp Lys Tyr Met Ala Asp Ser Val Leu Trp His Leu Lys

260 265 270

Asn Pro Gln Ser Glu Gln Lys Val Ile Val Val Ala His Asn Ala His

275 280 285

Ile Gln Lys Thr Pro Ile Leu Tyr Asp Gly Phe Leu Ser Cys Leu Pro

290 295 300

Met Gly Gln Arg Leu Lys Asn Ala Ile Gly Asp Asp Tyr Met Ser Leu

305 310 315 320

Gly Ile Thr Ser Tyr Ser Gly His Thr Ala Ala Leu Tyr Pro Glu Val

325 330 335

Asp Thr Lys Tyr Gly Phe Arg Val Asp Asn Phe Gln Leu Gln Glu Pro

340 345 350

Asn Glu Gly Ser Val Glu Lys Ala Ile Ser Gly Cys Gly Val Thr Asn

355 360 365

Ser Phe Val Phe Phe Arg Asn Ile Pro Glu Asp Leu Gln Ser Ile Pro

370 375 380

Asn Met Ile Arg Phe Asp Ser Ile Tyr Met Lys Ala Glu Leu Glu Lys

385 390 395 400

Ala Phe Asp Gly Ile Phe Gln Ile Glu Lys Ser Ser Val Ser Glu Val

405 410 415

Val Tyr Glu

<210> 86

<211> 388

<212> PRT

<213> Bacteroides fragilis (Bacteroides fragilis)

<400> 86

Met Thr Met Arg Ile Asp Thr Asp Lys Gln Met Asn Leu Leu Ser Asp

1 5 10 15

Lys Asn Val Ala Ile Ile Gly Gly Gly Pro Val Gly Leu Thr Met Ala

20 25 30

Lys Leu Leu Gln Gln Asn Gly Ile Asp Val Ser Val Tyr Glu Arg Asp

35 40 45

Asn Asp Arg Glu Ala Arg Ile Phe Gly Gly Thr Leu Asp Leu His Lys

50 55 60

Gly Ser Gly Gln Glu Ala Met Lys Lys Ala Gly Leu Leu Gln Thr Tyr

65 70 75 80

Tyr Asp Leu Ala Leu Pro Met Gly Val Asn Ile Ala Asp Lys Lys Gly

85 90 95

Asn Ile Leu Ser Thr Lys Asn Val Lys Pro Glu Asn Arg Phe Asp Asn

100 105 110

Pro Glu Ile Asn Arg Asn Asp Leu Arg Ala Ile Leu Leu Asn Ser Leu

115 120 125

Glu Asn Asp Thr Val Ile Trp Asp Arg Lys Leu Val Met Leu Glu Pro

130 135 140

Gly Lys Lys Lys Trp Thr Leu Thr Phe Glu Asn Lys Pro Ser Glu Thr

145 150 155 160

Ala Asp Leu Val Ile Leu Ala Asn Gly Gly Met Ser Lys Val Arg Lys

165 170 175

Phe Val Thr Asp Thr Glu Val Glu Glu Thr Gly Thr Phe Asn Ile Gln

180 185 190

Ala Asp Ile His Gln Pro Glu Ile Asn Cys Pro Gly Phe Phe Gln Leu

195 200 205

Cys Asn Gly Asn Arg Leu Met Ala Ser His Gln Gly Asn Leu Leu Phe

210 215 220

Ala Asn Pro Asn Asn Asn Gly Ala Leu His Phe Gly Ile Ser Phe Lys

225 230 235 240

Thr Pro Asp Glu Trp Lys Asn Gln Thr Gln Val Asp Phe Gln Asn Arg

245 250 255

Asn Ser Val Val Asp Phe Leu Leu Lys Glu Phe Ser Asp Trp Asp Glu

260 265 270

Arg Tyr Lys Glu Leu Ile His Thr Thr Leu Ser Phe Val Gly Leu Ala

275 280 285

Thr Arg Ile Phe Pro Leu Glu Lys Pro Trp Lys Ser Lys Arg Pro Leu

290 295 300

Pro Ile Thr Met Ile Gly Asp Ala Ala His Leu Met Pro Pro Phe Ala

305 310 315 320

Gly Gln Gly Val Asn Ser Gly Leu Val Asp Ala Leu Ile Leu Ser Asp

325 330 335

Asn Leu Ala Asp Gly Lys Phe Asn Ser Ile Glu Glu Ala Val Lys Asn

340 345 350

Tyr Glu Gln Gln Met Phe Met Tyr Gly Lys Glu Ala Gln Glu Glu Ser

355 360 365

Thr Gln Asn Glu Ile Glu Met Phe Lys Pro Asp Phe Thr Phe Gln Gln

370 375 380

Leu Leu Asn Val

385

<210> 87

<211> 267

<212> PRT

<213> Enterococcus faecium (Enterococcus faecium)

<400> 87

Met Leu Lys Gln Lys Glu Leu Ile Ala Asn Val Lys Asn Leu Thr Glu

1 5 10 15

Ser Asp Glu Arg Ile Thr Ala Cys Met Met Tyr Gly Ser Phe Thr Lys

20 25 30

Gly Glu Gly Asp Gln Tyr Ser Asp Ile Glu Phe Tyr Ile Phe Leu Lys

35 40 45

His Ser Ile Thr Ser Asn Phe Asp Ser Ser Asn Trp Leu Phe Asp Val

50 55 60

Ala Pro Tyr Leu Met Leu Tyr Lys Asn Glu Tyr Gly Thr Glu Val Val

65 70 75 80

Ile Phe Asp Asn Leu Ile Arg Gly Glu Phe His Phe Leu Ser Glu Lys

85 90 95

Asp Met Asn Ile Ile Pro Ser Phe Lys Asp Ser Gly Tyr Ile Pro Asp

100 105 110

Thr Lys Ala Met Leu Ile Tyr Asp Glu Thr Gly Gln Leu Glu Asn Tyr

115 120 125

Leu Ser Glu Ile Ser Gly Ala Arg Pro Asn Arg Leu Thr Glu Glu Asn

130 135 140

Ala Asn Phe Leu Leu Cys Asn Phe Ser Asn Leu Trp Leu Met Gly Ile

145 150 155 160

Asn Val Leu Lys Arg Gly Glu Tyr Ala Arg Ser Leu Glu Leu Leu Ser

165 170 175

Gln Leu Gln Lys Asn Thr Leu Gln Leu Ile Arg Met Ala Glu Lys Asn

180 185 190

Ala Asp Asn Trp Leu Asn Met Ser Lys Asn Leu Glu Lys Glu Ile Ser

195 200 205

Leu Glu Asn Tyr Lys Lys Phe Ala Lys Thr Thr Ala Arg Leu Asp Lys

210 215 220

Val Glu Leu Phe Glu Ala Tyr Lys Asn Ser Leu Leu Leu Val Met Asp

225 230 235 240

Leu Gln Ser His Leu Ile Glu Gln Tyr Asn Leu Lys Val Thr His Asp

245 250 255

Ile Leu Glu Arg Leu Leu Asn Tyr Ile Ser Glu

260 265

<210> 88

<211> 532

<212> PRT

<213> Artificial

<220>

<223> TEM36-GGGGGG-CTXM16

<400> 88

His Pro Glu Thr Leu Val Lys Val Lys Asp Ala Glu Asp Gln Leu Gly

1 5 10 15

Ala Arg Val Gly Tyr Ile Glu Leu Asp Leu Asn Ser Gly Lys Ile Leu

20 25 30

Glu Ser Phe Arg Pro Glu Glu Arg Phe Pro Met Val Ser Thr Phe Lys

35 40 45

Val Leu Leu Cys Gly Ala Val Leu Ser Arg Val Asp Ala Gly Gln Glu

50 55 60

Gln Leu Gly Arg Arg Ile His Tyr Ser Gln Asn Asp Leu Val Glu Tyr

65 70 75 80

Ser Pro Val Thr Glu Lys His Leu Thr Asp Gly Met Thr Val Arg Glu

85 90 95

Leu Cys Ser Ala Ala Ile Thr Met Ser Asp Asn Thr Ala Ala Asn Leu

100 105 110

Leu Leu Thr Thr Ile Gly Gly Pro Lys Glu Leu Thr Ala Phe Leu His

115 120 125

Asn Met Gly Asp His Val Thr Arg Leu Asp Arg Trp Glu Pro Glu Leu

130 135 140

Asn Glu Ala Ile Pro Asn Asp Glu Arg Asp Thr Thr Met Pro Ala Ala

145 150 155 160

Met Ala Thr Thr Leu Arg Lys Leu Leu Thr Gly Glu Leu Leu Thr Leu

165 170 175

Ala Ser Arg Gln Gln Leu Ile Asp Trp Met Glu Ala Asp Lys Val Ala

180 185 190

Gly Pro Leu Leu Arg Ser Ala Leu Pro Ala Gly Trp Phe Ile Ala Asp

195 200 205

Lys Ser Gly Ala Gly Glu Arg Gly Ser Arg Gly Ile Ile Ala Ala Leu

210 215 220

Gly Pro Asp Gly Lys Pro Ser Arg Ile Val Val Ile Tyr Thr Thr Gly

225 230 235 240

Ser Gln Ala Thr Met Asp Glu Arg Asp Arg Gln Ile Ala Glu Ile Gly

245 250 255

Ala Ser Leu Ile Lys His Trp Gly Gly Gly Gly Gly Gly Gln Thr Ser

260 265 270

Ala Val Gln Gln Lys Leu Ala Ala Leu Glu Lys Ser Ser Gly Gly Arg

275 280 285

Leu Gly Val Ala Leu Ile Asp Thr Ala Asp Asn Thr Gln Val Leu Tyr

290 295 300

Arg Gly Asp Glu Arg Phe Pro Met Cys Ser Thr Ser Lys Val Met Ala

305 310 315 320

Ala Ala Ala Val Leu Lys Gln Ser Glu Thr Gln Lys Gln Leu Leu Asn

325 330 335

Gln Pro Val Glu Ile Lys Pro Ala Asp Leu Val Asn Tyr Asn Pro Ile

340 345 350

Ala Glu Lys His Val Asn Gly Thr Met Thr Leu Ala Glu Leu Ser Ala

355 360 365

Ala Ala Leu Gln Tyr Ser Asp Asn Thr Ala Met Asn Lys Leu Ile Ala

370 375 380

Gln Leu Gly Gly Pro Gly Gly Val Thr Ala Phe Ala Arg Ala Ile Gly

385 390 395 400

Asp Glu Thr Phe Arg Leu Asp Arg Thr Glu Pro Thr Leu Asn Thr Ala

405 410 415

Ile Pro Gly Asp Pro Arg Asp Thr Thr Thr Pro Arg Ala Met Ala Gln

420 425 430

Thr Leu Arg Gln Leu Thr Leu Gly His Ala Leu Gly Glu Thr Gln Arg

435 440 445

Ala Gln Leu Val Thr Trp Leu Lys Gly Asn Thr Thr Gly Ala Ala Ser

450 455 460

Ile Arg Ala Gly Leu Pro Thr Ser Trp Thr Ala Gly Asp Lys Thr Gly

465 470 475 480

Ser Gly Gly Tyr Gly Thr Thr Asn Asp Ile Ala Val Ile Trp Pro Gln

485 490 495

Gly Arg Ala Pro Leu Val Leu Val Thr Tyr Phe Thr Gln Pro Gln Gln

500 505 510

Asn Ala Glu Ser Arg Arg Asp Val Leu Ala Ser Ala Ala Arg Ile Ile

515 520 525

Ala Glu Gly Leu

530

<210> 89

<211> 532

<212> PRT

<213> Artificial

<220>

<223> fusion CTXM16-6G-TEM36

<400> 89

Gln Thr Ser Ala Val Gln Gln Lys Leu Ala Ala Leu Glu Lys Ser Ser

1 5 10 15

Gly Gly Arg Leu Gly Val Ala Leu Ile Asp Thr Ala Asp Asn Thr Gln

20 25 30

Val Leu Tyr Arg Gly Asp Glu Arg Phe Pro Met Cys Ser Thr Ser Lys

35 40 45

Val Met Ala Ala Ala Ala Val Leu Lys Gln Ser Glu Thr Gln Lys Gln

50 55 60

Leu Leu Asn Gln Pro Val Glu Ile Lys Pro Ala Asp Leu Val Asn Tyr

65 70 75 80

Asn Pro Ile Ala Glu Lys His Val Asn Gly Thr Met Thr Leu Ala Glu

85 90 95

Leu Ser Ala Ala Ala Leu Gln Tyr Ser Asp Asn Thr Ala Met Asn Lys

100 105 110

Leu Ile Ala Gln Leu Gly Gly Pro Gly Gly Val Thr Ala Phe Ala Arg

115 120 125

Ala Ile Gly Asp Glu Thr Phe Arg Leu Asp Arg Thr Glu Pro Thr Leu

130 135 140

Asn Thr Ala Ile Pro Gly Asp Pro Arg Asp Thr Thr Thr Pro Arg Ala

145 150 155 160

Met Ala Gln Thr Leu Arg Gln Leu Thr Leu Gly His Ala Leu Gly Glu

165 170 175

Thr Gln Arg Ala Gln Leu Val Thr Trp Leu Lys Gly Asn Thr Thr Gly

180 185 190

Ala Ala Ser Ile Arg Ala Gly Leu Pro Thr Ser Trp Thr Ala Gly Asp

195 200 205

Lys Thr Gly Ser Gly Gly Tyr Gly Thr Thr Asn Asp Ile Ala Val Ile

210 215 220

Trp Pro Gln Gly Arg Ala Pro Leu Val Leu Val Thr Tyr Phe Thr Gln

225 230 235 240

Pro Gln Gln Asn Ala Glu Ser Arg Arg Asp Val Leu Ala Ser Ala Ala

245 250 255

Arg Ile Ile Ala Glu Gly Leu Gly Gly Gly Gly Gly Gly His Pro Glu

260 265 270

Thr Leu Val Lys Val Lys Asp Ala Glu Asp Gln Leu Gly Ala Arg Val

275 280 285

Gly Tyr Ile Glu Leu Asp Leu Asn Ser Gly Lys Ile Leu Glu Ser Phe

290 295 300

Arg Pro Glu Glu Arg Phe Pro Met Val Ser Thr Phe Lys Val Leu Leu

305 310 315 320

Cys Gly Ala Val Leu Ser Arg Val Asp Ala Gly Gln Glu Gln Leu Gly

325 330 335

Arg Arg Ile His Tyr Ser Gln Asn Asp Leu Val Glu Tyr Ser Pro Val

340 345 350

Thr Glu Lys His Leu Thr Asp Gly Met Thr Val Arg Glu Leu Cys Ser

355 360 365

Ala Ala Ile Thr Met Ser Asp Asn Thr Ala Ala Asn Leu Leu Leu Thr

370 375 380

Thr Ile Gly Gly Pro Lys Glu Leu Thr Ala Phe Leu His Asn Met Gly

385 390 395 400

Asp His Val Thr Arg Leu Asp Arg Trp Glu Pro Glu Leu Asn Glu Ala

405 410 415

Ile Pro Asn Asp Glu Arg Asp Thr Thr Met Pro Ala Ala Met Ala Thr

420 425 430

Thr Leu Arg Lys Leu Leu Thr Gly Glu Leu Leu Thr Leu Ala Ser Arg

435 440 445

Gln Gln Leu Ile Asp Trp Met Glu Ala Asp Lys Val Ala Gly Pro Leu

450 455 460

Leu Arg Ser Ala Leu Pro Ala Gly Trp Phe Ile Ala Asp Lys Ser Gly

465 470 475 480

Ala Gly Glu Arg Gly Ser Arg Gly Ile Ile Ala Ala Leu Gly Pro Asp

485 490 495

Gly Lys Pro Ser Arg Ile Val Val Ile Tyr Thr Thr Gly Ser Gln Ala

500 505 510

Thr Met Asp Glu Arg Asp Arg Gln Ile Ala Glu Ile Gly Ala Ser Leu

515 520 525

Ile Lys His Trp

530

<210> 90

<211> 537

<212> PRT

<213> Artificial

<220>

<223> fusion TEM36-G (EAAAK)2-CTXM16

<400> 90

His Pro Glu Thr Leu Val Lys Val Lys Asp Ala Glu Asp Gln Leu Gly

1 5 10 15

Ala Arg Val Gly Tyr Ile Glu Leu Asp Leu Asn Ser Gly Lys Ile Leu

20 25 30

Glu Ser Phe Arg Pro Glu Glu Arg Phe Pro Met Val Ser Thr Phe Lys

35 40 45

Val Leu Leu Cys Gly Ala Val Leu Ser Arg Val Asp Ala Gly Gln Glu

50 55 60

Gln Leu Gly Arg Arg Ile His Tyr Ser Gln Asn Asp Leu Val Glu Tyr

65 70 75 80

Ser Pro Val Thr Glu Lys His Leu Thr Asp Gly Met Thr Val Arg Glu

85 90 95

Leu Cys Ser Ala Ala Ile Thr Met Ser Asp Asn Thr Ala Ala Asn Leu

100 105 110

Leu Leu Thr Thr Ile Gly Gly Pro Lys Glu Leu Thr Ala Phe Leu His

115 120 125

Asn Met Gly Asp His Val Thr Arg Leu Asp Arg Trp Glu Pro Glu Leu

130 135 140

Asn Glu Ala Ile Pro Asn Asp Glu Arg Asp Thr Thr Met Pro Ala Ala

145 150 155 160

Met Ala Thr Thr Leu Arg Lys Leu Leu Thr Gly Glu Leu Leu Thr Leu

165 170 175

Ala Ser Arg Gln Gln Leu Ile Asp Trp Met Glu Ala Asp Lys Val Ala

180 185 190

Gly Pro Leu Leu Arg Ser Ala Leu Pro Ala Gly Trp Phe Ile Ala Asp

195 200 205

Lys Ser Gly Ala Gly Glu Arg Gly Ser Arg Gly Ile Ile Ala Ala Leu

210 215 220

Gly Pro Asp Gly Lys Pro Ser Arg Ile Val Val Ile Tyr Thr Thr Gly

225 230 235 240

Ser Gln Ala Thr Met Asp Glu Arg Asp Arg Gln Ile Ala Glu Ile Gly

245 250 255

Ala Ser Leu Ile Lys His Trp Gly Glu Ala Ala Ala Lys Glu Ala Ala

260 265 270

Ala Lys Gln Thr Ser Ala Val Gln Gln Lys Leu Ala Ala Leu Glu Lys

275 280 285

Ser Ser Gly Gly Arg Leu Gly Val Ala Leu Ile Asp Thr Ala Asp Asn

290 295 300

Thr Gln Val Leu Tyr Arg Gly Asp Glu Arg Phe Pro Met Cys Ser Thr

305 310 315 320

Ser Lys Val Met Ala Ala Ala Ala Val Leu Lys Gln Ser Glu Thr Gln

325 330 335

Lys Gln Leu Leu Asn Gln Pro Val Glu Ile Lys Pro Ala Asp Leu Val

340 345 350

Asn Tyr Asn Pro Ile Ala Glu Lys His Val Asn Gly Thr Met Thr Leu

355 360 365

Ala Glu Leu Ser Ala Ala Ala Leu Gln Tyr Ser Asp Asn Thr Ala Met

370 375 380

Asn Lys Leu Ile Ala Gln Leu Gly Gly Pro Gly Gly Val Thr Ala Phe

385 390 395 400

Ala Arg Ala Ile Gly Asp Glu Thr Phe Arg Leu Asp Arg Thr Glu Pro

405 410 415

Thr Leu Asn Thr Ala Ile Pro Gly Asp Pro Arg Asp Thr Thr Thr Pro

420 425 430

Arg Ala Met Ala Gln Thr Leu Arg Gln Leu Thr Leu Gly His Ala Leu

435 440 445

Gly Glu Thr Gln Arg Ala Gln Leu Val Thr Trp Leu Lys Gly Asn Thr

450 455 460

Thr Gly Ala Ala Ser Ile Arg Ala Gly Leu Pro Thr Ser Trp Thr Ala

465 470 475 480

Gly Asp Lys Thr Gly Ser Gly Gly Tyr Gly Thr Thr Asn Asp Ile Ala

485 490 495

Val Ile Trp Pro Gln Gly Arg Ala Pro Leu Val Leu Val Thr Tyr Phe

500 505 510

Thr Gln Pro Gln Gln Asn Ala Glu Ser Arg Arg Asp Val Leu Ala Ser

515 520 525

Ala Ala Arg Ile Ile Ala Glu Gly Leu

530 535