阅读说明：本技术 可溶性肾素(原)受体用于治疗代谢紊乱及相关疾患的用途 (Soluble feritin (original) receptor is used to treat the purposes of metabolic disorder and related disorders ) 是由 T·杨 于 2018-03-14 设计创作，主要内容包括：公开了治疗肥胖症或肥胖症相关疾患的方法,所述方法包括向肥胖或患有肥胖症相关疾患的受试者施用有效量的可溶性肾素(原)受体(sPRR)。在一些情况下,肥胖症相关疾患可以是但不限于脂肪变性、高血糖症、胰岛素抵抗、慢性肾病。公开了减轻体重的方法,所述方法包括向有需要的受试者施用有效量的sPRR。公开了治疗受试者的脂肪肝的方法,所述方法包括向有需要的受试者施用有效量的sPRR。公开了治疗体液和电解质紊乱的方法,所述方法包括向诊断为患有体液和电解质紊乱的受试者施用有效量的sPRR。(The method for the treatment of obesity or obesity related disorders is disclosed, the method includes to obesity or a effective amount of soluble feritin (original) receptor (sPRR) of the application of the subject with obesity related disorders.In some cases, obesity related disorders can be but not limited to steatosis, hyperglycemia, insulin resistance, chronic kidney disease.The method to lose weight is disclosed, the method includes applying a effective amount of sPRR to subject in need.The method of the fatty liver for the treatment of subject is disclosed, the method includes applying a effective amount of sPRR to subject in need.The method for the treatment of body fluid and electrolyte disturbance is disclosed, the method includes applying a effective amount of sPRR to the subject for being diagnosed as suffering from body fluid and electrolyte disturbance.)

1. a kind of method for treating body fluid and electrolyte disturbance, the method includes to being diagnosed as with body fluid and electrolyte disturbance A effective amount of soluble feritin (original) receptor (sPRR) of subject's application.

2. the method as described in claim 1, wherein the body fluid and electrolyte are central diabetes insipidus (CDI), kidney originality urine Flooding (NDI), haemorrhagic shock, septic shock or the decrement of the plasma volume due to caused by the various causes of disease.

3. such as method claimed in claims 1-2, the method also includes before applying the sPRR to the subject into The step of whether row test suffers from body fluid and electrolyte disturbance with the determination subject.

4. a kind of method for treating obesity or obesity related disorders, the method includes to obesity or with obesity correlation A effective amount of soluble feritin (original) receptor (sPRR) of subject's application of illness.

5. method as claimed in claim 4, wherein the obesity related disorders are steatosis, hyperglycemia, insulin Resistance or chronic kidney disease.

6. a kind of method to lose weight, the method includes applying a effective amount of soluble feritin to subject in need (original) receptor (sPRR).

7. a kind of method for the fatty liver for treating subject, the method includes to subject in need application is a effective amount of can Dissolubility feritin (original) receptor (sPRR).

8. such as method of any of claims 1-7, wherein applying a effective amount of sPRR includes that application carries effective quantity SPRR medium.

9. method according to claim 8, wherein the medium is nano particle.

10. method as claimed in claim 9, wherein the nano particle is liposome or polymer.

11. such as method of any of claims 1-10, wherein the sPRR is fusion protein.

12. the method as described in claim 1, the method also includes applying known therapeutic agent.

13. method as claimed in claim 12, wherein the known therapeutic agent becomes known for treating body fluid and electrolyte is disorderly Disorderly.

14. method as claimed in claim 4, the method also includes applying known therapeutic agent.

15. method as claimed in claim 14, wherein the known therapeutic agent becomes known for treating obesity or obesity Related disorders.

16. the method as described in claim 12 or 14, wherein the known therapeutic agent becomes known for treating fatty liver.

17. the method as described in claim 12 or 14, wherein the known therapeutic agent becomes known for losing weight.

18. the method as described in any one of claim 1-17, wherein the sPRR is wild type sPRR.

19. the method as described in any one of claim 1-17, wherein sPRR is identical as wild type sPRR 70%-99%.

20. the method as described in any one of claim 1-19, wherein the sPRR is applied with the amount of 10-300 μ g/kg/ days.

21. a kind of method for treating metabolic disorder, the method includes tested with body fluid and electrolyte disturbance to being diagnosed as A effective amount of soluble feritin (original) receptor (sPRR) of person's application.

22. method as claimed in claim 21, wherein the metabolic disorder be obesity, obesity related disorders, body fluid and Electrolyte disturbance, lipid disorders, diabetes, motion relevance hyponatremia, metabolic myopathy, pancreatitis, systemic fatty group Knit scorching, X linksystem hypophosphatemia.

Background technique

In the past twenty years, overweight and overweight people illness rate steeply rises.Currently, there is 65% adult in the U.S. It is overweight, there are 31% Obesity Among Adultss.Obesity is the master of diabetes B, steatosis, chronic kidney disease and other chronic diseases Want risk factor.Therefore, there are 18,000,000 Americans with diabetes, there is another American to be diagnosed with this per minute Kind disease.Available therapy is limited the control of obesity and obesity related disorders.Particularly, these therapies are most Number is directed to each component part of metabolic syndrome.There is an urgent need to develop a kind of whole or most of keys for targeting this disease The New therapies of component part.Thiazolidinedione (TZD), such as Rosiglitazone and Pioglitazone are the PPAR γ activation of synthesis Agent, they are highly effective to control of hyperglycemia since its insulin sensitizing agent is acted on, but it has the beneficial effect of lipodogramme Limit, and can even obesity be made to deteriorate.TZD and oedema, weight gain and the increase of the incidence of chronic heart failure etc. are main Want side effect associated.Need to relate generally to multiple component parts of metabolic syndrome, including obesity, height in intervening measure Blood glucose disease, steatosis and chronic kidney disease.

Summary of the invention

Overall length feritin (original) receptor (PRR) is the transmembrane receptor with 350 amino acid of feritin original and feritin, can make it It is subjected to the cracking of proteases mediate, is generated by N-terminal ectodomain, soluble feritin (original) receptor (sPRR) and 8.9kDa C-terminal The 28kDa albumen of intracellular domain (referred to as " M8.9 ") composition.This document describes the intervening measures based on sPRR, relate generally to generation Thank multiple component parts of syndrome, including obesity, hyperglycemia, steatosis and chronic kidney disease.

Disclosed herein is treatment obesity or the methods of obesity related disorders, and the method includes to obesity or with fertilizer A effective amount of soluble feritin (original) receptor (sPRR) of subject's application of fat disease related disorders.In some cases, obesity Related disorders can be but not limited to steatosis, hyperglycemia, insulin resistance, chronic kidney disease.

Disclosed herein is the methods to lose weight, a effective amount of the method includes applying to subject in need sPRR。

Disclosed herein is the methods of the fatty liver for the treatment of subject, and the method includes having to subject in need application The sPRR of effect amount.

Disclosed herein is the method for the treatment of body fluid and electrolyte disturbance, the method includes to being diagnosed as with body fluid and electricity The subject for solving matter disorder applies a effective amount of sPRR.

Disclosed herein is the method for the treatment of metabolic disorder, described includes to the subject's application being diagnosed as with metabolic disorder A effective amount of sPRR.

In some cases, sPRR can be fusion protein.In some cases, sPRR can be wild type sPRR.In Under some cases, sPRR is identical as wild type sPRR 70%-99%.

The other advantage of disclosed method and composition will be set forth in part in the description which follows, and from described It will partly understand in part, or can be known by practicing disclosed method and composition.Disclosed method And the advantages of composition by by being particularly pointed out in appended claims element and combination be achieved and obtained.It should be appreciated that Foregoing general description and subsequent detailed description are all only exemplary and illustrative, rather than to claimed The limitation of invention.

Detailed description of the invention

It is incorporated to this specification and the attached drawing for constituting this specification a part shows the several of disclosed method and composition A embodiment, and together with specification it is used to explain the principle of disclosed method and composition.

Figure 1A-Fig. 1 M shows the not isolabeling of the antibody of the different structure territory for PRR.(A-C) in continuous Rat renal Immunostaining is carried out on dirty slice, uses the anti-PRR-N antibody (amplification of the N-terminal structural domain (such as sPRR sequence) of identification PRR Rate: in A for 200 ×；It is 400 × in B), and identify the anti-PRR-C antibody (C of the C-terminal structural domain of PRR；Magnifying power: 400 times). (D-F) PRR antibody is incubated for altogether together with anti-AQP2 antibody.(G-I) merge image.Arrow in B and E indicates chief cell；C and Arrow in F indicates sandwich cell.Anti- PRR-N antibody label is primarily targeted for the top film of chief cell, in contrast anti-PRR-C Antibody label is primarily targeted for sandwich cell.(J-M) it is the specificity of verification mark, uses sPRR (J), the sPRR- with expression His (K) or its immune peptide (L) do not carry out immunostaining with the anti-PRR-N antibody of primary antibody (M) preincubate.Shown image represents Every group of three to six animals.

Fig. 2A -2E shows the characterization to sPRR function.PRR, that is, sPRR-His is recombinated in mammalian cell expression system In be expressed as fusion protein, the fusion protein includes the secreting signal peptide of sPRR, the histidine tag of C-terminal and N-terminal.(A) from It is purified into sPRR-His in culture medium, single 29.6kDa band is shown as in 12%SDS/PAGE gel.E1-E4 is to wash The sequence fraction of abjection.The Primary rat IMCD cell grown in Transwell is exposed to 10nM sPRR-His by (B and C) Continue 24 hours, and passes through quantitative RT-PCR and immunoblotting assay AQP2 expression (every group of n=5).(D) it is individually testing In, the CD cell of culture AQP2- Luciferase construct is transfected, it is grown to and converges, then uses medium or 10nM SPRR-His is handled 24 hours, and measures uciferase activity (every group of n=5).(E) STRING is for predicting protein-egg The algorithm based on template of white matter structure.Use the protein of STRING identification and sPRR interaction.The analysis discloses 10 A hit results: CTSG (cathepsin G), ACE2 (hypertensin 1 invertase 2), CMA1 (rotten enzyme 1), MME (film metal Endopeptidase), ACE (hypertensin 1 invertase 1), CTSZ (Ctsz), ENPEP (glutamy aminopeptidase), FZD8 (curling family receptors 8), WNT3A (no wing MMTV integrates 3A) and LRP6 (LDH receptor related protein 6). CTR, reference material.

Fig. 3 A-3D shows kidney FZD8 expression and its interaction with sPRR.(A) to mutual between sPRR and FZD8 The co-immunoprecipitation of effect is analyzed.The film fraction of rat kidney medulla is precipitated with anti-PRR-N antibody mediated immunity, and is examined with anti-FZD8 antibody It surveys, vice versa.(B) FZD8's under low magnifying power in rat kidney is immune labeled.(C) under high magnification in medullary substance outer layer FDZ8, AQP2 and NKCC2 it is immune labeled.The total label to AQP2 is carried out on same slice, it is enterprising being continuously sliced Label of the row to NKCC2.CD, concetrated pipe；PT, proximal tubule；TL, the thick section of ascending branch.(D) FDZ8 in rat medullary substance internal layer and PRR's is immune labeled.Serial section is dyed with anti-FZD8 antibody and anti-PRR-N antibody.It is marked altogether with anti-AQP2 antibody same Slice.Shown image represents every group of three to six animals.

Fig. 4 A-4E, which is shown, adjusts in AQP2 expression Wnt/ beta-catenin approach in response to sPRR-His processing Interaction in vitro analysis.(A) FZD8 caused by siRNA strikes low verifying (every group of n=3).Turn with or without FZD8 siRNA Contaminate Primary rat IMCD cell.FZD8 protein expression is assessed by immunoblotting.(B) presence or absence of FZD8siRNA's In the case of, influence (every group n=12) of the sPRR-His to Wnt response uciferase activity.It is transfected with or without FZD8 siRNA IMCD cell is handled 24 hours with the sPRR-His of 10nM later.Wnt/ beta-catenin is assessed using Cignal reporting system The activity of approach, data are indicated with relative response ratio.(C-E) FZD8 siRNA transfected into rat IMCD cell is used, with OMP or XAV- 939 pretreatments 1 hour, and handled 24 hours with 10nM sPRR.AQP2 protein expression is determined by immunoblotting assay.Density Variable is shown in below trace.(C) FZD8 strikes the influence (every group of n=6) of the low AQP2 protein expression to sPRR induction.(D) Influence (every group n=6) of the OMP to the sPRR-His AQP2 protein expression induced.(E) the AQP2 table that XAV induces sPRR-His The influence (every group of n=6) reached.Data are shown as average value ± SE；Compared to reference material, * P < 0.05；Compared to sPRR- His, #P < 0.05.

Fig. 5 A-5F shows Wnt/ beta-catenin approach in Primary rat IMCD cell to the acute and chronic of AVP Unique effect in response.IMCD cell XAV is pre-processed 1 hour, and is handled 30 minutes or 24 hours with 10nM AVP. (A) when AVP is handled 30 minutes, the cAMP (every group of n=6) in ELISA measurement culture medium is used.(B and C) is (every to film fraction Group n=6) (B) and cytoplasm fraction (every group of n=6) (C) progress AQP2 immunoblotting assay.(D-F) at 24 hours, measurement training It supports base cAMP (every group of n=6) (D), AQP2 immunoblotting assay (every group of n=6) (E) is carried out to full cell pyrolysis liquid, to total serum IgE It carries out AQP2mRNA quantitative RT PCR analysis (every group of n=4) (F).Data are shown as average value ± SE；Compared to reference material, * P < 0.05；Compared to independent AVP, #P < 0.05.

Fig. 6 A-6H shows effect of the Wnt/ beta-catenin approach in the urine concentrating capacity of rat.To SD rat Medium, OMP or XAV are applied, is placed in metabolic cage to evaluate under basal status (A-C) or the water after WD 24 hours (D-H) Metabolism state (every group of n=5 rat).Under basal status, measurement water intake (A), urine volume (B) and osmotic pressure of urine (C).(D- F) during 24 hours WD, monitoring urine volume (D), osmotic pressure of urine (E) and weight (BW) variation (F).(G and H) at the end of experiment, Measure plasma osmolarity (G) and Hct (H).Compared to reference material, * P < 0.05；Compared to only WD, #P < 0.05.

Fig. 7 A-7F shows Wnt/ beta-catenin approach and is adjusting kidney AQP2 expression during antidiuresis in rats In internal effect.(A and B) is in order to evaluate WD to the activation of kidney beta-catenin, to from control group and dehydration rat Cortex renis (every group of n=4 rat) (A) and medullary substance internal layer (every group of n=4 rat) (B) nuclear leve part carry out beta-catenin White immunoblotting assay.(C-F) it XAV (E) or is not added XAV (F) to adding OMP (C) with medium or WD or OMP (D) is not added or adds The cortex renis and medullary substance internal layer of the rat of processing carry out the analysis of AQP2 gene experssion (every group of n=5 rat).Data are shown For average value ± SE；Compared to reference material, * P < 0.05；Compared to WD, #P < 0.05.CO, cortex；IM, medullary substance internal layer.

Fig. 8 A- Fig. 8 E shows antidiuretic activity of the sPRR-His in mouse NDI model.It is led via implantation is jugular SPRR-His injection male C57/BL6 mouse is continued 7 days by pipe, and medium or V2R antagonist OPC31260 is then administered orally (OPC) continue 3 days.Mouse is placed in metabolic cage, to evaluate daily water intake and urinary output.(A) daily (every group of water intake N=4 mouse).(B) daily urinary output (every group of n=4 mouse).(C) osmotic pressure of urine (every group of n=4 mouse).(D) it urinates SPRR excretion (every group of n=4 mouse).Data are shown as average value ± SE.In A-C, compared to individual OPC, #P < 0.05；D, compared to reference material, * P < 0.05.(E) schematic diagram of sPRR mechanism of action.In distal nephron inner cavity, sPRR In conjunction with the FZD8 in the receptor complex on the film of chief cell top, beta-catenin is caused to activate, so that cAMP is promoted to accumulate, AQP2 transcription is eventually led to increase and urine concentration enhancing.

Fig. 9 A-9I shows initial characteristics of the TO901317 in vivo with external diuresis and PRR inhibiting effect.To Male C57/BL7 Mouse oral applies TO901317, and i.v. is transfused sPRR-His, continues 7 days.Receive medium processing Mouse as a control group.At the end of experiment, the collection of twenty-four-hour urine liquid is carried out, sPRR excretion is urinated by elisa assay later, And it is expressed by immunoblotting assay kidney PRR.(A) urinary output (every group of n=30 mouse).(B) osmotic pressure of urine (every group of n= 15 mouse).(C) immunoblotting assay (every group of n=15 mouse) of kidney PRR expression.(D) to the close of the immunoblotting in C Spend quantitative analysis.(E) to the elisa assay (every group of n=8 mouse) of urine sPRR.(F-I) CD cell of the TO901317 to culture In PRR expression influence.MpkCCD cell is exposed to medium or 10 μM of TO901317 continue 24 hours.Cell is split It solves liquid and carries out PRR protein expression immunoblotting assay, and using the concentration of sPRR in ELISA measurement culture medium, and pass through albumen Matter content is standardized.In individual experiment, cell PRR- Luciferase construct is transfected, it is made to grow to remittance It closes, is then handled 24 hours with medium or 10 μM of TO901317.(F) to the immunoblotting assay of PRR (every group of n=15). (G) densitometry of the immunoblotting in A is analyzed.(H) to the elisa assay of culture medium sPRR (every group of n=10).(I) glimmering Light element enzymatic determination (every group of n=5).

Figure 10 A-10E shows influence of the sPRR-His to the DI induced of TO901317 in mouse.Individually with medium or TO901317 or with sPRR-His Combined Treatment male C57/BL6 mouse, continue 7 days.(A) (every group of n=8 is only small for urinary output Mouse).(B) water intake (every group of n=8 mouse).(C) water surplus (every group of n=8 mouse).(D) osmotic pressure of urine (every group of n=8 Mouse).(E) Hct (every group of n=8 mouse).

Figure 11 A-11E shows the shadow of kidney AQP2 expression and urine feritin in the mouse that sPRR-His handles TO901317 It rings.Individually with medium or TO901317 or with sPRR-His Combined Treatment male C57/BL6 mouse, continue 7 days.By immune Trace and immunostained for analysis AQP2 expression.By the generation of measurement AngI to measure urine renin activity, and surveyed by ELISA Surely feritin original/renin concentration is urinated.(A) to the immunoblotting assay (every group of n=15 mouse) of AQP2 expression.(B) exempt in A The densitometry of epidemic disease trace is analyzed.(C) renin activity (every group of n=8 mouse) is urinated.(D) it is (every to urinate feritin original/feritin excretion N=8 mouse of group).(E) lxr agonist TO091317 (TO) inhibition PRR transcribes and induces the schematic diagram of the mechanism of DI.With The LXR that TO091317 is combined serves as the transition repressor of PRR gene, leads to the reduction of PRR/sPRR level, so that AQP2 table Up to reduction, DI is eventually led to.

Figure 12 shows influence of the sPRR-His to the weight of the male C57/BL6 mouse in the obesity of diet induced. Since 1 month big, male C57/BL6 is placed in high fat diet.During last 2 weeks, mouse is grouped at random to receive Medium or sPRR-His.Show the changes of weight in 2 weekly treatment periods.Every group of N=9.Data are average value ± SE.

Figure 13 shows influence of the sPRR-His to the perirenal fat of DIO mouse.

Figure 14 A and Figure 14 B show influence of the sPRR-His to the plasma glucose (A) and insulin (B) of DOI mouse. Blood glucose is measured by using glucose meter.By using EIA plasma insulin.Every group of N=9.Data are average value ± SE.

Figure 15 A and Figure 15 B show glucose tolerance test (GTT) and insulin tolerance test (ITT).Fasting 6 hours Afterwards, the glucose or insulin (0.25U/kg weight) of application single dose are injected via IP.A series of blood collections are carried out later And blood glucose measurement.Every group of N=9.Data are average value ± SE.

Figure 16 A- Figure 16 C shows influence of the sPRR-His to the steatosis of DIO mouse.(A) general appearance of liver. Show presentation graphics.(B) after being centrifuged in liver homogenate lipid layer appearance.It shows from every group of 4 representativenesses The sample of animal.(C) liver triglyceride content.Every group of N=9.Data are average value ± SE.

Influence Figure 17 shows sPRR-His to the plasma triglyceride of DIO mouse.Every group of N=9.Data are average Value ± SE.

Figure 18 shows influence of the sPRR-His to the albuminuria of DIO mouse.Urinary albumin is measured by EIA.Every group of N= 9.Data are average value ± SE.

Figure 19 A, Figure 19 B and Figure 19 C show sPRR-His to urine volume and epididymal adipose tissues amount and in the mouse of lithium processing In influence.Although the diuresis (Figure 19 A) for failing to influence Li induction is co-administered with sPRR-His, unexpectedly reduce Fat mass (Figure 19 B).The image of epididymal adipose tissues is shown in Figure 19 C.In other two groups of fat ratio in Li+sPRR-His group Fat is smaller, and more " brown ".Every group of N=4-5.Data are average value ± SE.

Specific embodiment

By reference to below to specific embodiment and wherein the detailed description of included example and attached drawing and its it Disclosed method and composition can be more easily to understand in preceding and description later.

It should be appreciated that unless otherwise specified, disclosed method and composition is not limited to specific synthetic method, spy Fixed analytical technology or specific reagent, therefore disclosed method and composition can change.It should also be understood that used herein Term is only used for the purpose of description specific embodiment, it is not intended that is limited.

Disclosed material, composition and component can be used for disclosed method and composition, can be with disclosed side Method and composition are used in combination, and can be used to the product for preparing disclosed composition or disclosed method and composition. Disclosed herein is these materials and other materials, it should be understood that if disclose the combination of these materials, subset, interaction, Group etc., without clearly disclosing specifically referring to for the arrangement to various individuals and collective combinations and these compounds, then Each case obtains clearly covering and describing for this paper.If disclosing molecule A, B and C and molecule D, E and F And the combined example of molecule A-D is disclosed, even if then also considered without each combination of independent narration individually and collectively Each combination.Therefore, in this example, if specifically considered in combination A-E, A-F, B-D, B-E, B-F, C-D, C-E and C-F Each, then should be considered as disclosing A, B and C；D, E and F；With example combination A-D.Equally, these any subset or Combination is also specifically considered and openly.It, should be by thus, for example, if specifically considering the subgroup of A-E, B-F and C-E It is considered as and discloses A, B and C；D, E and F；With example combination A-D.The concept is suitable for all aspects of the application, including but unlimited Step in the method for making and using disclosed composition.Therefore, it if can be carried out there are many additional step, answers Work as understanding, each of these additional steps can be with any specific embodiment of disclosed method or the group of embodiment Progress is unified, and has been combined as every kind and specifically considered and should be considered as obtaining disclosure.

A. it defines

It should be appreciated that disclosed method and composition is not limited to described ad hoc approach, scheme and reagent, because this A little disclosed method and compositions can change.It will also be appreciated that terms used herein are specific just for the sake of describing Embodiment, and be not intended to limit the scope of the invention, the scope of the present invention will be limited only by the appended claims.

It must be noted that unless context clearly dictates otherwise, otherwise as herein and used in the appended claims Singular "one", "an" and " described " include plural referents.Thus, for example, refer to " sPRR " include it is multiple in this way SPRR, refer to that " sPRR " is to refer to one or more sPRR and its equivalent known to those skilled in the art, etc..

" optional " or " optionally " indicates that the event, situation or the material that then describe may occur or exist or may not Occur or be not present, and the description includes the event, situation or material generation or existing example and the event, feelings The example that condition or material do not occur or be not present.

As used herein, term " subject " or " patient " refer to for example for experimental, diagnostic and/or therapeutic mesh , any organism of composition of the invention can be applied to it.Typical subject includes animal (for example, mammal is all Such as non-human primate and the mankind；Birds；Domestic animal or farm-animals, such as cat, dog, sheep, goat, ox, Ma He Pig；Laboratory animal, such as mouse, rat and cavy；Rabbit；Fish；Reptile；Zoo animal and wild animal).In general, " subject " is animal, including mammal such as people and primate；Etc..

As used herein, term " treatment " refers to partly or completely direct release, improvement, mitigation, delay specified disease, illness And/or the breaking-out of illness；Inhibit or slow down the progress of specified disease, illness and/or illness；Reduce specified disease, illness and/or The severity of illness；And/or reduce one or more symptoms of specified disease, illness and/or illness or the incidence of feature. Treatment can be applied to the subject for the sign for not showing disease, illness and/or illness and/or be applied to and only show disease The subject of the early indication of disease, illness and/or illness, it is associated with the disease, illness and/or illness to reduce development Symptom risk.

Term " therapeutic ", which refers to, can treat disease or illness or can improve one kind associated with disease or illness Or treatment, therapy or the drug of a variety of symptoms.

" effective quantity " refers to the active agents for being enough that required physiologic result is realized in the individual for needing active agents Amount.Effective quantity can according to subject to be treated health and physical condition, the taxonomy category of subject to be treated, The preparation of sPRR is varied evaluation and other correlative factors of subject's medical conditions between subjects.

Herein, range can be expressed as from " about " particular value and/or to " about " another particular value.Work as expression When such a range, unless the context clearly indicates otherwise, otherwise also specifically consider and think to disclose from a particular value And/or the range to another particular value.It similarly, should when value is expressed as approximation by using antecedent " about " Understand that otherwise the particular value forms another embodiment specifically considered, the implementation unless the context clearly indicates otherwise Scheme should be considered as obtaining disclosure.It will be further understood that unless the context clearly indicates otherwise, otherwise the endpoint phase of each range Another endpoint is all important, and unrelated with other endpoints.Finally, it will be understood that unless context in addition It explicitly pointing out, the subrange of all single values and value that are otherwise included in clear scope of disclosure is also all specifically contemplated, And it should be considered as obtaining disclosure.Whether it specifically discloses under specific circumstances some or complete in these embodiments Portion, above content are applicable in.

Unless otherwise defined, otherwise all technical and scientific terms used herein has and disclosed method and combines The identical meaning of the meaning that the technical staff in field belonging to object is generally understood.Although implementing or testing method of the invention With any method and material similar or equivalent with those described herein method and material can be used when composition, but it is present Particularly useful method, device and material are described.It publication cited herein and quotes involved by these publications To material be clearly incorporated to by reference hereby.Any content herein be understood not to recognize the present invention due to It formerly invents and haves no right prior in this disclosure.Do not recognize that any bibliography constitutes the prior art.Bibliography is begged for By set forth the opinion of its author, and applicant retains and queries the cited accuracy of document and the right of correlation.It should be clear Understand to Chu, although many publications incorporated herein, this reference is not an admission that any in these documents Document constitutes a part of general knowledge known in this field.

This specification throughout the specification and claims, the modification of word " comprising " and the word is such as "comprising" and " containing " expression " including but not limited to ", and be not intended to exclude such as other additives, component, integer or step. Particularly, it is being set fourth as including in one or more steps or the method for operation, it is particularly contemplated that each step includes institute The content (unless the step includes a restrictive term such as " consist of ") listed, it means that each step is equal Other additives, component, integer or the step that do not list in the step for example are excluded unintentionally.

B. the treatment method of metabolic disorder and related disorders

The method for the treatment of metabolic disorder is disclosed, described includes applying a effective amount of sPRR to subject.Disclose treatment The method of metabolic disorder, described includes to a effective amount of sPRR of subject's application being diagnosed as with metabolic disorder.When different in vivo When normal chemical reaction changes normal metabolic process, metabolic disorder will occur.The symptom of metabolic disorder may include drowsiness, abdomen Bitterly, weight loss, jaundice and epilepsy.In some cases, metabolic disorder includes but is not limited to obesity disease related to obesity Trouble, body fluid and electrolyte disturbance, lipid disorders, diabetes, motion relevance hyponatremia, metabolic myopathy, pancreatitis, whole body Property pimelitis (pansteatitis), X linksystem hypophosphatemia and lysosomal storage disease.

C. the treatment method of obesity or obesity related disorders

The method for the treatment of obesity or obesity related disorders is disclosed, the method includes to obesity or with obesity A effective amount of soluble feritin (original) receptor (sPRR) of subject's application of related disorders.In some cases, obesity is related Illness can be but not limited to steatosis, hyperglycemia, insulin resistance, chronic kidney disease.

The method for the treatment of obesity or obesity related disorders is disclosed, the method includes to obesity or with obesity A effective amount of soluble feritin (original) receptor (sPRR) of subject's application of related disorders, the method also includes in application sPRR The subject is tested so that whether the determination subject is fat or the step of with obesity related disorders before.

The method to lose weight is disclosed, the method includes applying a effective amount of sPRR to subject in need.In Under some cases, subject in need is any subject for being diagnosed as needing to lose weight.

The method of the fatty liver for the treatment of subject is disclosed, the method includes applying effective quantity to subject in need SPRR.In some cases, subject in need is to be diagnosed to be any subject with fatty liver.In some cases Under, fatty liver can be non-alcohol fatty liver (NAFLD) or alcoholic liver disease (ALD).

In some cases, applying a effective amount of sPRR includes the medium that application carries a effective amount of sPRR.Some In the case of, medium can be nano particle.For example, nano particle can be liposome or polymer.In some cases, matchmaker Jie's object can have targeting moiety, wherein sPRR can be targeted interested specific cells or tissue by the targeting moiety.

The method for the treatment of obesity or obesity related disorders is disclosed, the method includes to obesity or with obesity The subject of related disorders applies a effective amount of sPRR, and the method also includes applying known therapeutic agent.In some cases, The known therapeutic agent becomes known for treating obesity or obesity related disorders.For example, for treating obesity or obesity The known treatment agent of disease related disorders can be but not limited to GLP-1 receptor stimulating agent (such as Saxenda (r)).In some feelings Under condition, sPRR and known therapeutic agent are administered simultaneously.In some cases, sPRR and known therapeutic agent sequential application.One In a little situations, sPRR and known therapeutic agent are prepared together.In some cases, sPRR and known therapeutic agent are separately formulated.

The method to lose weight is disclosed, the method includes applying a effective amount of sPRR, institute to subject in need The method of stating further includes applying known therapeutic agent.In some cases, it is known that the known therapeutic agent can reduce weight.Example Such as, the known treatment agent for losing weight can be but be not limited to use in the identical treatment agent for the treatment of obesity.In some feelings Under condition, sPRR and known therapeutic agent are administered simultaneously.In some cases, sPRR and known therapeutic agent sequential application.One In a little situations, sPRR and known therapeutic agent are prepared together.In some cases, sPRR and known therapeutic agent are separately formulated.

The method of the fatty liver for the treatment of subject is disclosed, the method includes applying effective quantity to subject in need SPRR, the method also includes applying known therapeutic agent.In some cases, the known therapeutic agent becomes known for controlling Treat fatty liver.For example, the known treatment agent for treating fatty liver can be but not limited to statins and thiazolidinedione. In some cases, sPRR and known therapeutic agent are administered simultaneously.In some cases, sPRR and known therapeutic agent are applied in succession With.In some cases, sPRR and known therapeutic agent are prepared together.In some cases, sPRR and known therapeutic agent point Open preparation.

In some cases, sPRR can be fusion protein.For example, sPRR fusion partner can be but not limited to a group ammonia Sour (His) label, glutathione-S-transferase (GST) label, c-Myc label, hemagglutinin (HA) label or myelin basic egg White (MBP) label.

In some cases, sPRR can be wild type sPRR.For example, wild type sPRR can be SEQ ID NO:1's Sequence.ANEFSILRSPGSVVFRNGNWPIPGDRIPDVAALSMGFSVKEDLSWPGLAVGNLFHRPRATIMVTVKGVDKL ALPTGSVISYPLENAVPFSLDSVANSIHSLFSEETPVVLQLAPSEERVYMVGKANSVFEDLSVTLRQLRNRLFQEN SVLNSLPLNSLSRNNEVDLLFLSELQVLHDISSLLSRHKHLAKDHSPDLYSLELAGLDELGKRYGEDSEQFRDASR ILVDALQKFADDMYSLYGGNAVVELVTVKSFDTSL (SEQ ID NO:1)

In some cases, sPRR is identical as wild type sPRR 70%-99%.In some cases, substituted amino acid It can be amino acid, conservative substitution or the non-conservative substitutions of modification.

In some cases, sPRR can be applied with the amount of 10-300 μ g/kg/ days.In some cases, dosage regimen can Including single administration sPRR.In some cases, dosage regimen may include once a week, twice a week, three-times-weekly, on every Thursdays Secondary, secondary on every Fridays, secondary on every Saturdays or seven application sPRR weekly, for 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 Week, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 Week, 26 weeks, 28 weeks, 30 weeks, 32 weeks, 34 weeks, 36 weeks, 38 weeks, 40 weeks, 42 weeks, 44 weeks, 46 weeks, 48 weeks, 50 weeks or 52 weeks.

D. the treatment method of body fluid and electrolyte disturbance

The method for the treatment of body fluid and electrolyte disturbance is disclosed, the method includes to being diagnosed as with body fluid and electrolyte The subject of disorder applies a effective amount of sPRR.In some cases, body fluid and electrolyte disturbance include but is not limited to central Diabetes insipidus (CDI), diabetes insipidus,nephrogenic (NDI), haemorrhagic shock, septic shock and the blood plasma due to caused by the various causes of disease hold Amount decrement (plasma volume contraction).

The method for the treatment of body fluid and electrolyte disturbance is disclosed, the method includes to being diagnosed as with body fluid and electrolyte The subject of disorder applies a effective amount of sPRR, and the method also includes testing before applying sPRR the subject The step of whether suffering from body fluid and electrolyte disturbance with the determination subject.

In some cases, applying a effective amount of sPRR includes the medium that application carries a effective amount of sPRR.Some In the case of, medium can be nano particle.For example, nano particle can be liposome or polymer.In some cases, matchmaker Jie's object can have targeting moiety, wherein sPRR can be targeted interested specific cells or tissue by the targeting moiety.

The method for the treatment of body fluid and electrolyte disturbance is disclosed, the method includes to being diagnosed as with body fluid and electrolyte The subject of disorder applies a effective amount of sPRR, and the method also includes applying known therapeutic agent.In some cases, described Known therapeutic agent becomes known for treating body fluid and electrolyte disturbance.For example, for treating the known of body fluid and electrolyte disturbance Therapy can be but not limited to thiazolidinedione, insulin, melbine, sugared diuretics (glycodiuretic).In some feelings Under condition, sPRR and known therapeutic agent are administered simultaneously.In some cases, sPRR and known therapeutic agent sequential application.One In a little situations, sPRR and known therapeutic agent are prepared together.In some cases, sPRR and known therapeutic agent are separately formulated.

In some cases, sPRR can be fusion protein.For example, sPRR fusion partner can be but not limited to a group ammonia Sour (His) label, glutathione-S-transferase (GST) label, c-Myc label, hemagglutinin (HA) label or myelin basic egg White (MBP) label.

In some cases, sPRR can be wild type sPRR.For example, wild type sPRR can be SEQ ID NO:1's Sequence.

In some cases, sPRR is identical as wild type sPRR 70%-99%.In some cases, substituted amino acid It can be amino acid, conservative substitution or the non-conservative substitutions of modification.

In some cases, sPRR can be applied with the amount of 10-300 μ g/kg/ days.In some cases, dosage regimen can Including single administration sPRR.In some cases, dosage regimen may include once a week, twice a week, three-times-weekly, on every Thursdays Secondary, secondary on every Fridays, secondary on every Saturdays or seven application sPRR weekly, for 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 Week, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 Week, 26 weeks, 28 weeks, 30 weeks, 32 weeks, 34 weeks, 36 weeks, 38 weeks, 40 weeks, 42 weeks, 44 weeks, 46 weeks, 48 weeks, 50 weeks or 52 weeks.

E. composition and preparation

Disclose the composition comprising the peptide containing sPRR.

As described herein, in some cases, sPRR can be wild type sPRR.For example, wild type sPRR can be SEQ The sequence of ID NO:1.In some cases, sPRR is identical as wild type sPRR 70%-99%.In some cases, substituted Amino acid can be amino acid, conservative substitution or the non-conservative substitutions of modification.

In some cases, sPRR can be fusion protein.For example, sPRR fusion partner can be but not limited to a group ammonia Sour (His) label, glutathione-S-transferase (GST) label, c-Myc label, hemagglutinin (HA) label or myelin basic egg White (MBP) label.

In some cases, it can pharmaceutically be prepared in acceptable carrier or together with pharmaceutically acceptable carrier And/or application sPRR.As used herein, term " pharmaceutically acceptable carrier " refer to sterile aqueous or non-aqueous solution, Dispersion liquid, suspension or lotion, and for being reconstructed into the sterile of sterile Injectable solution or dispersion liquid immediately before use Powder.Suitable aqueous and non-aqueous supporting agent, diluent, solvent or medium example include water, ethyl alcohol, polyalcohol (such as Glycerol, propylene glycol, polyethylene glycol etc.), carboxymethyl cellulose and its suitable mixture, vegetable oil (olive oil) and injectable Organic ester such as ethyl oleate.For example, (just being divided by using coating material such as lecithin by the granularity needed for maintaining For dispersion liquid) and by using surfactant, mobility appropriate can be maintained.These compositions can also contain adjuvant, Such as preservative, wetting agent, emulsifier and dispersing agent.It can be by the inclusion of various antibacterial agents and antifungal agent such as para hydroxybenzene Formic acid esters, chlorobutanol, phenol, sorbic acid etc. ensure to prevent the effect of microorganism.It it may also be desirable to comprising isotonic agent, such as Sugar, sodium chloride etc..Injectable drug form can be extended by the inclusion of the agent such as aluminum monostearate and gelatin that delay absorbs Absorption.It can be by forming drug in biodegradable polymer such as polylactide-polyglycolide, poly- (ortho esters) and gathering Microcapsule matrix in (acid anhydrides) is made the depot forms of injectable.According to the ratio of drug and polymer and used specific poly- The property for closing object, can control the rate of release of drug.It can also be by the way that drug to be trapped in the lipid compatible with bodily tissue Depot injectable formulations are prepared in body or microemulsion.It can be for example by being filtered via bacteria-retaining filter or by mixing The bactericidal agent for entering aseptic solid composite form sterilizes to injectable formulation, and the bactericidal agent can dissolve immediately before use Or it is dispersed in sterile water or other sterile injectable mediums.Suitable inert carrier may include sugared such as lactose.It is desirable that extremely The active ingredient particle of few 95 weight % has the effective size of grain in 0.01 to 10 micron range.

Therefore, the compositions disclosed herein may include lipid such as liposome, such as cationic-liposome (for example, DOTMA, DOPE, DC-cholesterol) or anionic liposome.If desired, liposome also may include protein in order to target spy Determine cell.Composition comprising peptide and cationic-liposome can be applied to blood, be applied to target organ, or be drawn into breathing To target respiratory tract cell in road.For example, can be by the group comprising peptide as described herein or nucleic acid sequence and cationic-liposome Close the pneumonocyte that object is applied to subject.About liposome, see, e.g., Brigham et al. Am.J.Resp.Cell.Mol.Biol.1:95-100 (1989)；Felgner et al. Proc.Natl.Acad.Sci USA 84: 7413-7417(1987)；United States Patent (USP) No.4,897,355.Furthermore, it is possible to using compound as particular cell types can be targeted The component of the microcapsules of such as macrophage is applied, or wherein the diffusion for special speed or dose design compound or Delivering of the compound from microcapsules.

On the one hand, disclose pharmaceutical composition, described pharmaceutical composition include any disclosed peptide as described herein or its Pharmaceutically acceptable salt or solvate and pharmaceutically acceptable carrier, diluent or diluent.In all fields, The peptide of pharmaceutical composition is encapsulated in delivery vehicle.On the other hand, delivery vehicle is liposome, microcapsules or nanometer Particle.On the other hand, delivery vehicle is PEGylated.

In method described herein, cell can be delivered the composition to via number of mechanisms.As hereinbefore defined, originally Text discloses the composition comprising any one or more of peptide as described herein, nucleic acid, carrier and/or antibody, can be used for Composition is generated, the composition can also include such as pharmaceutically acceptable carrier of carrier.For example, disclosing comprising herein The pharmaceutical composition of disclosed peptide and pharmaceutically acceptable carrier.On the one hand, it discloses comprising disclosed compound Pharmaceutical composition.I.e., it is possible to provide at least one disclosed compound comprising therapeutically effective amount or by disclosed side The pharmaceutical composition of at least one product and pharmaceutically acceptable carrier that method generates.

In some aspects, disclosed pharmaceutical composition include as active constituent disclosed compound (including its Pharmaceutically acceptable one or more salt), pharmaceutically acceptable carrier and optional other therapeutic ingredient or adjuvant.This The composition of invention includes being suitable for those of oral, rectum, topical and parenteral (including subcutaneous, intramuscular and intravenous) application group Object is closed, but most suitable approach will be targeted depending on specific host, applied active constituent in any given situation The property and severity of illness.Pharmaceutical composition exists in which can be convenient with unit dosage forms, and can pass through pharmaceutical field It is prepared by well known any method.

In all fields, pharmaceutical composition is disclosed, described pharmaceutical composition includes pharmaceutically acceptable carrier or dilute The disclosed compound for releasing agent and the therapeutically effective amount as active constituent, the production generated by disclosed preparation method Object, its pharmaceutically acceptable salt, solvate or polymorph, its hydrate, its solvate, its polymorph or it is vertical Body chemical isomer form.On the other hand, disclosed compound, product, its medicine for being generated by disclosed preparation method Acceptable salt, solvate or polymorph, its hydrate, its solvate, its polymorph or its spatial chemistry on Isomeric forms or its any subgroup or combination can be formulated into various medicament forms for applying purpose.

As used herein, term " pharmaceutically acceptable salt " refer to by pharmaceutically acceptable nontoxic alkali or acid preparation Salt.When the compound of the present invention is acid, corresponding salt can be convenient by pharmaceutically acceptable nontoxic alkali (including nothing Machine alkali and organic base) it prepares.Salt derived from such inorganic base includes aluminium salt, ammonium salt, calcium salt, copper (copper and cuprous) salt, iron Salt, ferrous salt, lithium salts, magnesium salts, manganese (manganese and sub- manganese) salt, sylvite, sodium salt, zinc salt etc..Particularly preferably ammonium salt, calcium salt, magnesium Salt, sylvite and sodium salt.Salt derived from pharmaceutically acceptable organic nontoxic alkali includes primary amine, secondary amine and tertiary amine and cyclammonium The salt of such as both naturally occurring and synthetic substituted amine with substituted amine.Pharmaceutically acceptable can have with other of forming salt The nontoxic alkali of machine includes ion exchange resin, such as arginine, glycine betaine, caffeine, choline, N, N '-dibenzyl-ethylenediamin, Diethylamine, 2- DEAE diethylaminoethanol, 2-dimethylaminoethanol, ethanol amine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, Aminoglucose (glucamine), histidine, breathes out amine (hydrabamine), isopropylamine, relies aminoglucose amine (glucosamine) Propylhomoserin, methylglucosamine, morpholine, piperazine, piperidines, polyamino resin, procaine, purine, theobromine, triethylamine, trimethylamine, three Propylamine, tromethamine etc..

As used herein, term " pharmaceutically acceptable non-toxic acid " includes inorganic acid, organic acid and salt prepared therefrom, Such as acetic acid, benzene sulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, salt Acid, Yi Xi sulfuric acid (isethionic), lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid (pamoic), pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-methyl benzenesulfonic acid etc..Preferably citric acid, hydrobromic acid, salt Acid, maleic acid, phosphoric acid, sulfuric acid and tartaric acid.

For therapeutic use, the salt of disclosed compound is that wherein ion balance is those of pharmaceutically acceptable Salt.But it can also be used for the system of for example pharmaceutically acceptable compound by the salt that bronsted lowry acids and bases bronsted lowry acceptable in non-pharmaceutical is formed In standby or purifying.All salt, regardless of whether be it is pharmaceutically acceptable, be included within the scope of the invention.

Mean to include disclosed compound in the pharmaceutically acceptable bronsted lowry acids and bases bronsted lowry addition salts referred to above or below The non-toxic acid and addition salt forms with therapeutic activity being capable of forming.Pharmaceutically acceptable acid-addition salts can be convenient ground It is obtained and with this suitable acid processing alkali form.Acid appropriate includes such as inorganic acid, such as halogen acids (such as hydrochloric acid Or hydrobromic acid), sulfuric acid, nitric acid, phosphoric acid etc.；Or organic acid, such as acetic acid, propionic acid, hydroxyacetic acid, lactic acid, pyruvic acid, grass Sour (i.e. ethanedioic acid), malonic acid, succinic acid (i.e. succinic acid), maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, methylsulphur Acid, ethanesulfonic acid, benzene sulfonic acid, p-methyl benzenesulfonic acid, cyclamic acid (cyclamic), salicylic acid, PAS, pamoic acid etc.. On the contrary, can be by converting free alkali form for the salt form with alkali process appropriate.

The disclosed compound containing acid proton can also turn and with organic base appropriate and inorganic alkali process Turn to its nontoxic metal or amine addition salt form.Base salt forms appropriate include: such as ammonium salt, alkali metal salt and alkaline-earth metal Salt, such as lithium, sodium, potassium, magnesium, calcium salt etc.；The salt formed by organic base, such as primary, secondary and tertiary aliphatic series and aromatic amine, such as first Amine, ethamine, propylamine, isopropylamine, four kinds of butylamine isomers, dimethylamine, diethylamine, diethanol amine, di-n-propylamine, diisopropylamine, two N-butylamine, pyrrolidines, piperidines, morpholine, trimethylamine, triethylamine, tripropyl amine (TPA), quinuclidine, pyridine, quinoline and isoquinolin, benzyl star are green Mycin, breathes out amine salt at N- methyl-D-glucosamine；And by the amino acids formed salt such as such as arginine, lysine.On the contrary Ground, can be by converting free acid form for salt form with acid processing.

In practice, the compounds of this invention of the invention or its pharmaceutically acceptable salt can match according to conventional drug Mixed technology is closely mixed as active constituent with pharmaceutical carriers.Carrier can take many forms, this is depended on needed for application Dosage form, such as oral or extra-parenteral (including intravenous).Therefore, pharmaceutical composition of the invention can be used as such as capsule The discrete unit for being suitable for being administered orally of agent, cachet or tablet exists, and each unit is containing the active constituent of predetermined amount.This Outside, composition can be with pulvis, granule, solution, the suspension in waterborne liquid, non-aqueous liquid, oil-in-water emulsion or oil The form of packet aqueous precursor emulsion exists.Other than general formulation listed above, the compound of the present invention and/or its pharmaceutically Acceptable one or more salt can also be applied by controlled release component and/or delivery apparatus.The composition can be by appointing What method of pharmacy preparation.In general, such method includes making active constituent in conjunction with the carrier for constituting one or more neccessary compositions The step of.In general, by mixing uniformly and closely active constituent with liquid carrier or the solid carriers of subdivision or both It closes to prepare composition.Then product can be easily shaped to required appearance.

In order to apply convenient and dose uniformity, foregoing pharmaceutical composition is configured to unit dosage forms and is particularly advantageous.Such as Unit dosage forms used herein refer to the physically discrete unit for being suitable as unit dose, and each unit contains that be computed can Combine the active constituent of the predetermined amount of therapeutic effect needed for generating with required pharmaceutical carriers.The example of such unit dosage forms is tablet (tablet including indentation or coating), capsule, pill, pulvis parcel, wafer (wafer), suppository, Injectable solution Or suspension etc. and its composite dosage form separated (segregatedmultiples).

Therefore, pharmaceutical composition of the invention may include pharmaceutically acceptable carrier and the compound of the present invention or chemical combination The pharmaceutically acceptable salt of object.As it is well known to the skilled in the art, " pharmaceutically acceptable " refers to such material Or carrier, it will be selected such that any degradation of active constituent minimizes and makes any adverse side effect in subject most Smallization.The compound of the present invention or its pharmaceutically acceptable salt can also be with one or more other treatment reactive compound groups Ground is closed to be included in pharmaceutical composition.

Used pharmaceutical carriers can be such as solid, liquid or gas.The example of solid carriers includes lactose, gypsum Powder, sucrose, talcum powder, gelatin, agar, pectin, Arabic gum, magnesium stearate and stearic acid.The example of liquid carrier be syrup, Peanut oil, olive oil and water.The example of gaseous state carrier includes carbon dioxide and nitrogen.Other examples of carrier include two nutmegs Acyl phospholipids acyl (dimyristoylphosphatidyl, DMPC), phosphate buffered saline (PBS) or multivesicular liposome.For example, PG: PC: cholesterol: peptide or PC: peptide can be used as the carrier in the present invention.Other suitable pharmaceutically acceptable carriers and its preparation It is described in (the 19th edition) volume A.R.Gennaro of Remington:The Science and Practice of Pharmacy, Mack Publishing Company, Easton, PA 1995.In general, in the formulation using the pharmaceutically acceptable of appropriate amount Salt so that preparation is isotonic.Other examples of pharmaceutically acceptable carrier include but is not limited to salt water, Ringer's solution and the right side Revolve sugar juice.The pH value of solution can be about 5 to about 8, or about 7 to about 7.5.Other carrier includes sustained release preparation such as containing The semipermeable matrices of the solid hydrophobic polymers of composition are stated, the matrix is the form of molded article, such as film, bracket (in angioplasty in implantable intravascular), liposome or particle.It will be apparent to those skilled in the art that certain Carrier may be it is furthermore preferred that this depends on the concentration of such as administration method and applied composition.It is most typical in these to incite somebody to action It is the standard carrier for being applied to people, the solution including such as sterile water, salt water and the buffer solution of physiological ph.

In order to improve solubility and/or stability of the disclosed peptide in pharmaceutical composition, it may be possible to advantageously adopt With α-, β-or gamma-cyclodextrin or derivatives thereof, the especially cyclodextrin that replaces of hydroxyalkyl, such as 2-HP-BETA-CD or Sulfobutyl ether β _ cyclodextrin.It is molten in pharmaceutical composition that the cosolvent of such as alcohol can also improve compound according to the present invention Xie Du and/or stability.

Pharmaceutical composition also may include carrier, thickener, diluent, buffer, preservative etc., as long as not damaging the present invention Polypeptide, peptide, nucleic acid, carrier expection activity.Pharmaceutical composition also may include one or more active constituents (in addition to this Except the composition of invention), antimicrobial, anti-inflammatory agent, anesthetic etc..It depends on whether locally or systemically to treat And region to be treated is depended on, pharmaceutical composition can be applied in many ways.

Due to being easy to apply, oral administration is preferred, and tablets and capsules represent best oral dose list Position form, obviously uses solid drugs carrier in this case.In the composition for preparing peroral dosage form, it can be used any Convenient drug media.For example, water, glycol, oil, alcohol, flavoring agent, preservative, colorant etc. can be used to form oral solution system Agent such as suspension, elixir and solution；And such as starch, sugar, microcrystalline cellulose, diluent, granulation agent, lubricant, adhesive, The carrier of disintegrating agent etc. can be used to form oral solid formulation such as pulvis, capsule and tablet.Due to being easy to apply, tablet and Capsule is preferred oral dosage units, thus uses solid drugs carrier.Optionally, tablet can by standard aqueous or Nonaqueous techniques are coated.

Composition for oral administration includes pulvis or granule, the suspension in water or non-aqueous medium or molten Liquid, capsule, sachet or tablet.Thickener, flavoring agent, diluent, emulsifier, dispersing aid or adhesive may be needs 's.Some compositions may by it is pharmaceutically acceptable acid or base addition salts in the form of apply, the salt be by with such as salt Acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, the inorganic acid of sulfuric acid and phosphoric acid and such as formic acid, acetic acid, propionic acid, glycolic, Lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid and fumaric acid organic acid reaction and formed；Or by with Such as sodium hydroxide, ammonium hydroxide, the inorganic base of potassium hydroxide and such as mono-, di-, trialkyl and arylamine and substituted second The organic base of hydramine is reacted and is formed.

Tablet containing composition of the invention can be prepared by suppressing or moulding, and be optionally added into one or more auxiliary Co-ingredients or adjuvant.Compressed tablets can be by suppressing the free-flowing form of such as pulvis or granule in suitable machine Active constituent, optionally prepared with adhesive, lubricant, inert diluent, surfactant or dispersant.Molding Tablet can be prepared by the mixture for the powder compound that molding is soaked with inert liquid diluent in suitable machine.

Pharmaceutical composition of the invention include as active constituent peptide such as sPRR (or its pharmaceutically acceptable salt), Pharmaceutically acceptable carrier and optional one or more other therapeutic agents or adjuvant.Composition of the invention includes being suitable for mouth Those of clothes, rectum, topical and parenteral (including subcutaneous, intramuscular and intravenous) application composition, but in any given feelings Most suitable approach will depend on the property and severity of the targeted illness of specific host, applied active constituent under condition. Pharmaceutical composition exists in which can be convenient with unit dosage forms, and can be prepared by any method well-known in the field of pharmacy.

Pharmaceutical composition of the invention suitable for parenteral administration can be prepared into reactive compound solution in water or Suspension.It may include suitable surfactant, such as hydroxypropyl cellulose.Can also glycerol, liquid macrogol and Dispersion liquid is prepared in the oil in its mixture.Furthermore, it is possible to prevent the obnoxious growth of microorganism comprising preservative.

It include aseptic aqueous solution or dispersion liquid suitable for injecting the pharmaceutical composition of the invention used.In addition, described group This sterile injectable solution of extemporaneous preparation or dispersion liquid can be used in the form of aseptic powdery by closing object.In all cases, Final injectable forms must be sterile, and must effectively flow to be easy to inject.Pharmaceutical composition must made It makes and keeps stablizing under condition of storage；It is therefore preferred that carrying out anti-corrosion with the pollution of preventing microorganism such as bacterium and fungi.Carrier Can be containing such as water, ethyl alcohol, polyalcohol (such as glycerol, propylene glycol and liquid macrogol etc.), vegetable oil and they Suitable mixture solvent or decentralized medium.

For example, Injectable solution can be prepared, wherein carrier includes saline solution, glucose solution or saline solution and Portugal The mixture of grape sugar juice.Injectable suspensions can also be prepared, in which case it is possible to use liquid carrier appropriate, Suspending agent etc..It further include the Solid form preparations for being intended to be converted into liquid form preparation immediately before use.

The preparation of parenteral administration includes sterile aqueous or non-aqueous solution, suspension and lotion.The reality of non-aqueous solvent Example is the organic ester (such as ethyl oleate) of propylene glycol, polyethylene glycol, vegetable oil (such as olive oil) and injectable.Aqueous carrier Including water, alcohol/aqueous solution, lotion or suspension, including salt water and buffer medium.Parenteral vehicles include that sodium chloride is molten Liquid, woods grignard dextrose, dextrose and sodium chloride, Lactated Ringer'S Solution or fixed oil.Intravenous vehicles include body fluid With nutritional supplement, electrolyte replenisher (based on those of woods grignard dextrose) etc..There may also be preservatives and its His additive, such as antimicrobial, antioxidant, chelating agent and inert gas etc..

Pharmaceutical composition of the invention can be in the form of being suitable for local use, such as aerosol, cream, ointment Agent, lotion, face powder, mouthwash, gargle etc..In addition, the composition can be in the form of suitable for transdermal device.These Preparation can be prepared via conventional processing method using the compound of the present invention or its pharmaceutically acceptable salt.For example, By mixing hydrophilic material and water together with the compound of about 5wt% to about 10wt% to generate the emulsifiable paste with required consistency Agent or ointment, to prepare cream or ointment.

In the composition for being suitable for transdermal administration, carrier optionally includes penetration enhancers and/or suitable wetting agent, is appointed The suitable additive of any property of selection of land combination small percentage, the additive will not generate apparent harmful work to skin With.The additive can promote application to skin and/or can contribute to prepare required composition.These compositions can To apply in various ways, such as the application in the form of transdermal patch, point paint, ointment.

Pharmaceutical composition of the invention can be in the form of being suitable for rectal administration, and wherein carrier is solid.It is preferably mixed Close the suppository that object forms unit dose.Suitable carrier includes cupu oil and other materials commonly used in the art.Pass through head First the composition is mixed with the carrier being softened or melted, then cooling in a mold and molding, forms bolt with can be convenient Agent.

Formulations for topical administration may include ointment, lotion, cream, gelling agent, drops, suppository, spray, liquid Body and pulvis.Conventional pharmaceutical carrier (water base, powder base or oil base), thickener etc. can be required or needs.

In addition to aforementioned carrier ingredient, said medicine preparation can also optionally include one or more other carrier ingredients, Such as diluent, buffer, flavoring agent, adhesive, surfactant, thickener, lubricant, preservative (including antioxidant) Deng.In addition, may include other adjuvants so that preparation and the blood of expected recipient are isotonic.Containing the compound of the present invention and/or The composition of its pharmaceutically acceptable salt can also be prepared in the form of powder or liquid concentrate.

As it is well known to the skilled in the art, exact administration dosage and frequency depend on specifically disclosed compound, The product, its pharmaceutically acceptable salt, solvate or the polymorph that are formed by disclosed preparation method, its hydrate, Its solvate, its polymorph or its form of three-dimensional chemical isomer；The specific illness treated and the illness treated Severity；It is specific to the various factors of the medical history to the subject of its administration dosage, such as age；The body of particular subject The other drugs that weight, gender, illness degree and general physical condition and the individual may taken.Again it is apparent that The daily dose can be according to the reaction of the subject treated and/or commenting according to the doctor for issuing the compounds of this invention Valence and reduce or increase.

Depending on method of application, pharmaceutical composition will include 0.05 weight % to 99 weight %, preferably 0.1 weight % to 70 The active constituent of weight %, more preferable 0.1 weight % to 50 weight %；And 1 weight % to 99.95 weight %, preferably 30 weights The pharmaceutically acceptable carrier of % to 99.9 weight %, more preferable 50 weight % to 99.9 weight % is measured, all percentages are equal Total weight based on composition.

Positive allosteric is being needed to adjust in the active treatment condition of metabotropic glutamate receptor, suitable dosage level is usual For daily every kg patient's weight about 0.01 to 1000mg, and can be applied with single dose or multi-dose.In all fields, dosage Level would be about 0.1 to about 500mg/kg/ days, about 0.1 to 250mg/kg/ days or about 0.5 to 100mg/kg/ days.Suitable agent Amount level can be about 0.01 to 1000mg/kg/ days, about 0.01 to 500mg/kg/ days, about 0.01 to 250mg/kg/ days, about 0.05 to 100mg/kg/ days or about 0.1 to 50mg/kg/ days.In the range, dosage can be 0.05 to 0.5,0.5 to 5.0, Or 5.0 to 50mg/kg/ days.For being administered orally, preferably with comprising 1.0 to 1000 milligrams of active constituents, particularly 1.0,5.0, 10,15,20,25,50,75,100,150,200,250,300,400,500,600,750,800,900 and 1000 milligrams of activity at The form of the tablet divided provides the composition, so as to the dosage for adjusting patient to be treated of suiting the medicine to the illness.Can 1 time a day to 4 times, It is preferred that scheme once or twice daily applies the compound.This adjustable dosage regimen is anti-to provide optimal treatment It answers.

Can daily repeatedly, such as 2 times a day, 3 times, 4 times, 5 times or 6 application such units described above and below Dosage.In all fields, such unit dose can be applied 1 time or 2 times daily, so that the accumulated dose of 70kg adult is being applied every time In the range of every kg subject's weight 0.001 to about 15mg.On the other hand, each applied dose is every kg subject's body 0.01 to about 1.5mg is weighed, and this therapy can be with continued for several weeks or several months, and is in some cases the several years.But such as It is well-known to those skilled in the art, it should be understood that the specific dosage level of any particular patient will depend on many factors, packet The activity of particular compound used by including；Age, weight, general health situation, gender and the diet of the individual of patient； The time of application and approach；Excretion rate；Used other drugs are previously applied；And the serious journey of specified disease being treated Degree.

Typical dosage can be the 1mg repeatedly taken once a day or daily to about 100mg tablet or 1mg to about 300mg, or contain the release capsule or tablet of higher active component content once a day and in proportion.It can pass through Dissolved gum capsule material at various ph values, by the capsule by osmotic pressure slow release or by controlling known to any other Section loosen one's grip to obtain the effect of delay release.

The skilled person will be apparent that in some cases it may be necessary to other than using these ranges Dosage.Further, it is noted that clinician or attending physician combine the reaction of individual patient, it will know how and when open Begin, interrupt, adjust or terminate therapy.

The invention further relates to it is a kind of manufacture for adjust the glutamate receptor activity in mammal (such as people) (for example, Treat one or more neural and/or phrenoblabias associated with glutamic acid dysfunction) medicament method, the method Including one or more disclosed compounds, product or composition are mixed with pharmaceutically acceptable carrier or diluent. Therefore, on the one hand, the present invention relates to a kind of methods for manufacturing medicament, and the method includes will be at least one disclosed Compound or at least one disclosed product are mixed with pharmaceutically acceptable carrier or diluent.

Disclosed pharmaceutical composition also may include other treatment reactive compound, these therapeutical active compounds are usually used In the above-mentioned disease for the treatment of, illness or illness.

It should be appreciated that disclosed composition can be prepared by disclosed compound.It should also be understood that disclosed combination Object can be used in disclosed application method.

As already mentioned, the present invention relates to a kind of pharmaceutical composition, described pharmaceutical composition includes therapeutically effective amount Disclosed compound, product, its pharmaceutically acceptable salt, solvate or the polycrystalline generated by disclosed preparation method Type object, its hydrate, its solvate, its polymorph and pharmaceutically acceptable carrier.Moreover, it relates to one The method that kind prepares pharmaceutical composition, it is characterised in that according to the present invention by pharmaceutically acceptable carrier and therapeutically effective amount Compound closely mix.

As already mentioned, the invention further relates to a kind of pharmaceutical composition, described pharmaceutical composition include disclosed peptide, Its pharmaceutically acceptable salt, solvate or polymorph and one or more other drugs, the composition is for controlling Treatment, prevention, control, the disclosed compound of improvement or the other drugs may be to its effective disease or illness or reduction diseases Disease or illness risk；This composition is further related to for manufacturing the purposes of medicament.The invention further relates to disclosed peptides, its pharmacy The combination of upper acceptable salt, solvate or polymorph and anticancer therapeutic agent.Each in terms of other, the invention further relates to The combination of disclosed peptide, its pharmaceutically acceptable salt, solvate or polymorph.The invention further relates to be used as drug This combination.The different pharmaceutical of this combination or product can combine together with pharmaceutically acceptable carrier or diluent in list In a preparation or they can be present in individual preparation with pharmaceutically acceptable carrier or diluent.

In some cases, sPRR can be applied with the amount of 10-300 μ g/kg/ days.In some cases, dosage regimen can Including single administration sPRR.In some cases, dosage regimen may include once a week, twice a week, three-times-weekly, on every Thursdays Secondary, secondary on every Fridays, secondary on every Saturdays or seven application sPRR weekly, for 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 Week, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 Week, 26 weeks, 28 weeks, 30 weeks, 32 weeks, 34 weeks, 36 weeks, 38 weeks, 40 weeks, 42 weeks, 44 weeks, 46 weeks, 48 weeks, 50 weeks or 52 weeks.

F. it applies

In method described herein, can via number of mechanisms by sPRR or known therapeutic agent apply or be delivered to by Examination person.For example, sPRR or known therapeutic agent can be configured to pharmaceutical composition.

It depends on whether locally or systemically to treat and depend on region to be treated, can apply in many ways Pharmaceutical composition.

The preparation of parenteral administration includes sterile aqueous or non-aqueous solution, suspension and lotion.The reality of non-aqueous solvent Example is the organic ester (such as ethyl oleate) of propylene glycol, polyethylene glycol, vegetable oil (such as olive oil) and injectable.Aqueous carrier Including water, alcohol/aqueous solution, lotion or suspension, including salt water and buffer medium.Parenteral vehicles include that sodium chloride is molten Liquid, woods grignard dextrose, dextrose and sodium chloride, Lactated Ringer'S Solution or fixed oil.Intravenous vehicles include body fluid With nutritional supplement, electrolyte replenisher (based on those of woods grignard dextrose) etc..There may also be preservatives and its His additive, such as antimicrobial, antioxidant, chelating agent and inert gas etc..

Formulations for topical administration may include ointment, lotion, cream, gelling agent, drops, suppository, spray, liquid Body and pulvis.Conventional pharmaceutical carrier (water base, powder base or oil base), thickener etc. can be required or needs.

Composition for oral administration includes pulvis or granule, the suspension in water or non-aqueous medium or molten Liquid, capsule, sachet or tablet.Thickener, flavoring agent, diluent, emulsifier, dispersing aid or adhesive may be needs 's.Some compositions can by it is pharmaceutically acceptable acid or base addition salts in the form of apply, the salt be by with such as salt Acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, the inorganic acid of sulfuric acid and phosphoric acid and such as formic acid, acetic acid, propionic acid, glycolic, Lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid and fumaric acid organic acid reaction and formed；Or by with Such as sodium hydroxide, ammonium hydroxide, the inorganic base of potassium hydroxide and such as mono-, di-, trialkyl and arylamine and substituted second The organic base of hydramine is reacted and is formed.

G. medicine box

Materials described above and other materials can be packaged together in any suitable combination, be become for executing Or help to execute the medicine box of disclosed method.It is beneficial that the kit components in given medicine box by design, are suitble in institute's public affairs It is used together in the method opened.For example, disclosing disorderly for treating obesity, obesity related disorders and body fluid and electrolyte Random medicine box, the medicine box includes sPRR and the medium for carrying sPRR.The medicine box for treating metabolic disorder is disclosed, The medicine box includes sPRR and the medium for carrying sPRR.