阅读说明：本技术 通过鞘氨醇-1-磷酸途径增强癌症治疗功效 (Enhancing cancer treatment efficacy via sphingosine-1-phosphate pathway ) 是由 克里斯蒂安·奥克莱尔 于 2018-08-17 设计创作，主要内容包括：本发明涉及化疗之前的新辅助剂。本发明涉及选自鞘氨醇-1-磷酸裂解酶抑制剂的鞘氨醇-1-磷酸途径激活剂,其在化疗之前作为新辅助剂用于治疗癌症。鞘氨醇-1-磷酸途径激活剂通过瘤内血管网络的正常化而提高化疗功效,从而促进依次施用抗癌药或序贯放疗的效果。(The present invention relates to a novel adjuvant used before chemotherapy. The present invention relates to sphingosine-1-phosphate pathway activators selected from sphingosine-1-phosphate lyase inhibitors for use as neoadjuvant for the treatment of cancer prior to chemotherapy. Sphingosine-1-phosphate pathway activators enhance the efficacy of chemotherapy through normalization of the intratumoral vascular network, thereby promoting the effects of sequential administration of anticancer drugs or sequential radiation therapy.)

1. A compound for use in a method of treating cancer in a subject, wherein:

the compounds are sphingosine-1-phosphate lyase inhibitors that increase plasma concentrations of sphingosine-1-phosphate;

the compounds inhibit sprouting angiogenesis and stabilize blood vessels; and is

The compounds are novel adjuvants for enhancing the efficacy of sequential chemotherapy or radiotherapy.

2. The compound for use according to claim 1, wherein the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of 1-deoxy-dihydrosphingosine-1-phosphonic acid; 2-vinylsphinganine-1-phosphate; n- (4- (1H-imidazol-1-yl) benzyl) -4-amino-5-oxo-2- (piperidin-1-yl) -8- ((tetrahydrofuran-2-yl) methyl) -5, 8-dihydro-pyrido [2,3-d ] pyrimidine-6-carboxamide; (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine; (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methyl-piperazin-1-yl) benzonitrile; (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methyl-piperazin-1-yl) phthalazine-6-carbonitrile; 5- (hydroxymethyl) -2, 4-dimethyl-piperidin-3-ol; and

a compound of the formula (I),

wherein

R₁Represents H, acetyl, propionyl, N-hydroxyaminoacetyl, isoAzol-3-yl, 4, 5-dihydroisoOxazol-3-yl or prop-1-en-2-yl; said different isAzol-3-yl or 4, 5-dihydroisoOxazol-3-yl is optionally substituted with at least one alkyl, acetyl, haloalkyl, hydroxyalkyl, alkoxy, alkylamino, or nitro group;

R₂represents 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, ethyl carboxylate, N-methylformamide, 1, 2-dihydroxyethyl, 1-hydroxyethyl, 2-hydroxypropan-2-yl or a phosphate thereof; and is

R₃Represents H, methyl, ethyl, methoxy or alkoxyalkyl.

3. The compound for use according to claim 1 or 2, wherein the sphingosine-1-phosphate lyase inhibitor is a compound of formula (I) wherein:

R₁represents acetyl, N-hydroxyaminoacetyl or isoAzol-3-yl or 4, 5-dihydroisoOxazol-3-yl;

R₂represents a 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, 1, 2-dihydroxyethyl, 1-hydroxyethyl or 2-hydroxypropan-2-yl group; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

4. The compound for use according to any one of claims 1 to 3, wherein the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-2-acetyl-4 (5) -tetrahydroxybutyl-imidazole,

-1- (4- (1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime,

-1- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol,

-1- (4- (1-hydroxymethyl) -1H-imidazol-2-yl) ethanone,

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone oxime, and

- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) methanol.

5. The compound for use according to any one of claims 1 to 4, wherein the sphingosine-1-phosphate pathway activator is 2-acetyl-4 (5) -tetrahydroxybutyl-imidazole.

6. The compound for use according to any one of claims 1 to 5, wherein the sequential chemotherapy involves a single anticancer drug or a combination of anticancer drugs.

7. The compound for use according to any one of claims 1 to 6, wherein the sequential chemotherapy does not involve drugs showing anti-angiogenic properties.

8. The compound for use according to any one of claims 1 to 7, wherein the sequential chemotherapy involves a drug selected from gemcitabine, paclitaxel, cisplatin and oxaliplatin.

9. The compound for use according to any one of claims 1 to 8, wherein the subject exhibits a plasma concentration of sphingosine-1-phosphate that is less than the median plasma concentration of sphingosine-1-phosphate in healthy volunteers, said plasma concentrations being measured in the same experimental protocol in the subject and in healthy volunteers.

10. The compound for use according to any one of claims 1 to 9, wherein the subject exhibits a plasma concentration of sphingosine-1-phosphate of less than 700nM, preferably less than 650 nM.

11. The compound for use according to any one of claims 1 to 10, wherein the cancer is a solid tumor cancer.

12. The compound for use according to any one of claims 1 to 11, wherein the cancer is pancreatic cancer, preferably pancreatic cancer.

13. An in vitro non-invasive method for predicting the clinical outcome of a chemotherapy or radiotherapy treatment of a subject suffering from pancreatic cancer, wherein said method comprises:

a) detecting and quantifying the concentration of sphingosine-1-phosphate in a plasma sample previously obtained from a subject, an

b) Comparing the result obtained in step a) with a reference value; said reference value for sphingosine-1-phosphate is at least 10% or at least 15% lower than the median sphingosine-1-phosphate plasma concentration in healthy volunteers; wherein the plasma concentrations are measured in the same experimental protocol in the subject and in healthy volunteers.

14. The in vitro non-invasive method according to claim 13, wherein the reference value is 650nM sphingosine-1-phosphate.

15. A solid tumor cancer prognosis kit for performing the method according to claim 14, consisting of a set of reagents suitable for purifying and/or quantifying sphingosine-1-phosphate.

Technical Field

The present invention relates to the field of cancer treatment, such as chemotherapy and radiation therapy, and enhancing the efficacy of cancer treatment. More specifically, the present invention relates to sphingosine-1-phosphate pathway activators for use in methods of enhancing the efficacy of chemotherapy or radiation therapy. The compounds used in the methods of the invention, known as neoadjuvants, are administered to a subject prior to subsequent chemotherapy or radiotherapy. According to the invention, the novel adjuvant is a sphingosine-1-phosphate lyase inhibitor.

Background

Sphingosine-1-phosphate (S1P) is the product of sphingosine phosphorylation by sphingosine kinase. Like many products of sphingomyelin, it exhibits a wide range of biological activities and may subsequently be involved in various diseases, conditions or disorders.

S1P has pleiotropic effects on a variety of pathologies, including inflammation, diabetes, vascular disease (Bolick et al, Arterioscler Thromb Vasc Biol,2005,25, 976-.

Among the various possible effects, S1P enhances cell growth and inhibits the normal apoptotic response to various stimuli Manggau et al (J Clin Investig Dermatol,2001,117, pp 1241-1249) explain calcitriol (1 α, 25-dihydroxy vitamin D)₃) Protecting keratinocytes from apoptosis. One of the mechanisms of pharmacological action of calcitriol is the activation of sphingosine kinase activity, resulting in elevated levels of S1P.

There is substantial evidence that S1P is involved in cancer at various levels and involves the S1P receptor, sphingosine kinase, S1P phosphatase and S1P lyase. Wang et al (Mol cancer ther,2008,7,1993-2002) showed that S1P stimulates migration and invasion of malignant Epithelial Ovarian Cancer (EOC) cells, but inhibits migration of non-malignant Human Ovarian Surface Epithelial (HOSE) cells. Secondly, Riccitelli et al (Plos one,2013,8(6): e68229) explain the malignant nature of extracellular S1P in promoting Glioblastoma Stem Cells (GSCs), which are a subpopulation of cells within a tumor, resulting in resistance to temozolomide (temozolomide). Third, Kunkel et al (Nat Rev Drug Discov,2013,12,688-702) indicated that S1P, which is abnormally produced in ovarian cancer patients, is involved in the regulation of key cellular processes that promote initiation and progression of ovarian cancer, whereas drugs that block the S1P signaling pathway inhibit the growth of ovarian cancer cells. Kunkel et al further explain that S1P is known to form functional blood vessels.

However, Sultan et al (Journal of cancer,4,2013,315-319) disclose the synergistic effect of co-administration of 10 μ M of S1P with three chemotherapeutic drugs having anti-angiogenic properties, namely docetaxel, doxorubicin and cyclophosphamide. This document discloses that S1P increases the cytotoxicity of these drugs on breast cancer metastatic cell lines.

Meanwhile, WO03/097028 discloses a biphasic role of S1P receptor agonists in angiogenesis. On the one hand, high concentrations of S1P receptor agonists exhibit anti-angiogenic effects, which are useful in cancer therapy because they limit tumor angiogenesis. On the other hand, administration of a low concentration of S1P receptor agonist has an angiogenesis promoting effect. Promoting angiogenesis is useful for treating non-cancer related diseases such as wound healing, myocardial perfusion disorders, or vascular dysfunction associated with diabetes. According to the disclosure of WO03/097028, S1P receptor agonists may be used alone or in combination with one or more other anticancer drugs to produce a synergistic therapeutic effect. For example, an S1P receptor agonist may be used in combination with a chemotherapeutic agent for the treatment of lymphoid or myeloid cancers.

Despite the advances in pharmacology, the treatment of cancer has not been satisfactory. More particularly, there remains a need to take advantage of the angiogenic properties of S1P to normalize the vascular network within a tumor, thereby improving the efficacy of any anti-cancer drug that does not have anti-angiogenic properties. Thus, applicants have discovered a new approach to enhancing the efficacy of cancer therapy in the search for new therapeutic approaches to cancer therapy.

Contrary to the evidence of the prior art, inhibition of the S1P pathway leads to favorable regulation of key cellular processes contributing to tumor initiation and progression to cancer, applicants observed that treated patients with elevated plasma levels of S1P concentration showed better overall survival. Activation of S1P on S1P receptor 1 enhances the formation of adhesive junctions, inhibits vascular endothelial growth factor signaling, inhibits budding and stabilizes new vascular junctions.

The method according to the invention relates to the administration of sphingosine-1-phosphate pathway activators as neoadjuvant. The neoadjuvant according to the present invention inhibits sprouting angiogenesis and stabilizes blood vessels. Consequently, the chemotherapeutic agent subsequently has better accessibility to the tumor cells and therefore has an enhanced efficacy in its therapeutic effect. Also, the stabilization of the neovascular network improves the efficacy of sequential anticancer radiotherapy.

Disclosure of Invention

The present invention relates to a method for improving the efficacy of chemotherapy or radiotherapy, which comprises administering to a subject in need thereof a compound known as a neoadjuvant prior to chemotherapy or radiotherapy.

More particularly, the invention relates to compounds for use in methods of treating cancer in a subject. According to the invention, the compounds are sphingosine-1-phosphate lyase inhibitors which inhibit sprouting angiogenesis and stabilize blood vessels. The compounds used according to the invention are neoadjuvants which are used to enhance the efficacy of sequential chemotherapy.

The sphingosine-1-phosphate lyase inhibitor is selected from 1-deoxy-dihydrosphingosine-1-phosphonic acid; 2-vinylsphinganine-1-phosphate; n- (4- (1H-imidazol-1-yl) benzyl) -4-amino-5-oxo-2- (piperidin-1-yl) -8- ((tetrahydrofuran-2-yl) methyl) -5, 8-dihydropyrido [2,3-d ] pyrimidine-6-carboxamide; (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine; (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile; (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methyl-piperazin-1-yl) phthalazine-6-carbonitrile, 5- (hydroxymethyl) -2, 4-dimethyl-piperidin-3-ol, and compounds of formula (I)

Wherein

R₁Represents H, acetyl, propionyl, N-hydroxyaminoacetyl, isoAzol-3-yl, 4, 5-dihydroisoOxazol-3-yl or prop-1-en-2-yl; said different isAzol-3-yl or 4, 5-dihydroisoThe oxazol-3-yl is optionally substituted with at least one alkyl, acetyl, haloalkyl, hydroxyalkyl, alkoxy, alkylamino, or nitro group.

R₂Represents 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, ethyl carboxylate, N-methylformamide, 1, 2-dihydroxyethyl, 1-hydroxyethyl, 2-hydroxypropan-2-yl or a phosphate thereof; and is

R₃Represents H, methyl, ethyl, methoxy or alkoxyalkyl.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is a compound of formula (I) wherein:

R₁represents acetyl, N-hydroxyaminoacetyl or isoAzol-3-yl or 4, 5-dihydroisoAn azole-3-yl group, which is,

R₂represents a 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, 1, 2-dihydroxyethyl, 1-hydroxyethyl or 2-hydroxypropan-2-yl group; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of 2-acetyl-4 (5) -tetrahydroxybutyl-imidazole, 1- (4- (1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime, 1- (2- (iso-hydroxybutyl) -1H-imidazol-2-yl)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol, 1- (4- (1-hydroxymethyl) -1H-imidazol-2-yl) ethanone, 1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone oxime, and (2- (iso-methyl) -1H-imidazol-2-yl) ethanone oximeOxazol-3-yl) -1H-imidazol-4-yl) methanol.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is 2-acetyl-4 (5) -tetrahydroxybutyl-imidazole.

The sequential chemotherapy according to the present invention involves a single anticancer drug or a combination of anticancer drugs. Preferably, none of these drugs exhibit anti-angiogenic properties. In one embodiment, the sequential chemotherapy involves a drug selected from the group consisting of gemcitabine, paclitaxel, cisplatin, and oxaliplatin.

In one embodiment, the subject exhibits a plasma concentration of sphingosine-1-phosphate that is less than the median plasma concentration of sphingosine-1-phosphate in healthy volunteers. Since the measurement of plasma sphingosine-1-phosphate concentration depends on the analytical method, plasma concentrations were measured in the same experimental protocol in subjects and healthy volunteers.

In one embodiment, the subject exhibits a plasma concentration of sphingosine-1-phosphate of less than 700nM, preferably less than 650 nM.

In one embodiment, the cancer is a solid tumor cancer, preferably a pancreatic cancer, even more preferably a pancreatic adenocarcinoma.

In a second aspect, the present invention relates to an in vitro non-invasive method for predicting or monitoring the clinical outcome of a chemotherapy or radiotherapy treatment of a subject suffering from a solid tumor cancer, preferably pancreatic cancer. The in vitro non-invasive method according to the invention comprises the following steps:

a) detecting and quantifying the concentration of sphingosine-1-phosphate in a plasma sample previously obtained from the subject; and

b) comparing the result obtained in step a) with a reference value; the reference value for sphingosine-1-phosphate is at least 10% or at least 15% lower than the median sphingosine-1-phosphate plasma concentration in healthy volunteers; wherein the plasma concentrations are measured in the same experimental protocol in the subject and in healthy volunteers.

In one embodiment, the reference value for sphingosine-1-phosphate is 650 nM.

In a third aspect, the invention relates to a solid tumor cancer prognosis kit for performing the in vitro prognosis method of the invention, consisting of a set of reagents suitable for purifying and/or quantifying sphingosine-1-phosphate.

Definition of

In the present invention, the following terms have the following meanings:

- "alkyl" means a radical of the formula C_nH_2n+1Wherein n is a number greater than or equal to 1. Typically, the alkyl groups of the present invention contain 1 to 5 carbon atoms, preferably 1 to 4 carbon atoms. The alkyl group may be straight-chain or branched, and may have a substituent as described herein. Suitable alkyl groups include methyl, ethyl, propyl (n-propyl, isopropyl), butyl (n-butyl, isobutyl, sec-butyl and tert-butyl).

- "alkylamino" refers to monoalkylamino and dialkylamino groups represented by-NHR and-NRR ', where R and R' are alkyl groups. In one embodiment, R and R' are the same or different alkyl groups.

- "alkoxy" or "alkyloxy" means any alkyl-O-group.

- "alkoxyalkyl" means any alkyl-O-alkyl group. In one embodiment, alkoxymethyl refers to methoxymethyl (MOM).

"anti-angiogenesis" refers to natural substances, drugs or other compounds that can disrupt or interfere with the vascular micro-network required for tumor growth and metastasis.

"apoptosis" refers to programmed cell death.

An "agonist" is a molecule that interacts with and activates a membrane receptor.

"neoadjuvant" is a product administered before the main treatment, with the aim of enhancing the efficacy or increasing the effectiveness of the chemotherapy.

- "sphingosine-1-phosphate" or S1P is a signaling sphingolipid, also known as a lysosphingolipid. It is also known as a bioactive lipid mediator. Sphingolipids form a class of lipids characterized by specific aliphatic amino alcohols, which are sphingosines. Sphingosine kinase catalyzes sphingosine phosphate to S1P. As shown in FIG. 1, S1P can be dephosphorylated to sphingosine by sphingosine phosphatase. In addition, S1P can be degraded by the enzyme sphingosine phosphate lyase.

"tumorigenic" defines cells that can regress into tumors.

- "endothelium": endothelial cells are cells that line the inner surface of blood vessels. Thus, they are cells that come into direct contact with blood and ensure the integrity of blood vessels.

Drawings

FIG. 1 is a diagram showing the physiology of sphingosine-1-phosphate.

Fig. 2 includes a graph and two tables showing the plasma concentration of S1P at baseline in patients diagnosed with pancreatic cancer compared to healthy controls.

FIG. 3 is a table showing the median overall survival values for a subset of patients with "low" or "high" plasma concentrations of S1P (p-value: 0.05).

Figure 4 is a graph showing the risk ratio associated with overall survival as a function of S1P plasma concentration in patients treated with a conventional gemcitabine regimen.

Figure 5 is a comparative graph showing median overall survival for patients with S1P concentrations above 650nM and treated with gemcitabine or with the gemcitabine + kinase inhibitor masitinib combination.

Detailed Description

The present invention relates to compounds for use in a method of treating cancer in a subject, wherein the compounds are sphingosine-1-phosphate pathway activators, wherein the compounds used inhibit sprouting angiogenesis and stabilize blood vessels, and wherein the compounds used according to the invention are neoadjuvants for enhancing the efficacy of sequential chemotherapy or radiotherapy. In one embodiment, the compound used is a sphingosine-1-phosphate pathway activator, which enhances the plasma concentration of sphingosine-1-phosphate.

In one embodiment, the compounds used inhibit sprouting angiogenesis and stabilize blood vessels, and wherein the compounds used according to the invention are neoadjuvants for enhancing the efficacy of sequential chemotherapy.

Without wishing to be bound by theory, the effect of the neoadjuvant according to the invention leads to a stabilization of the neovascular perfusion of the tumor. Enhanced oxygenation of tumor cells enhances the efficacy of subsequent radiation therapy to a subject in need thereof.

In one embodiment, the compounds used inhibit sprouting angiogenesis and stabilize blood vessels, and wherein the compounds used according to the invention are neoadjuvants for enhancing the efficacy of sequential radiotherapy.

The plasma concentration value of sphingosine-1-phosphate can be determined according to the analytical method and sex.

In one embodiment, the subject exhibits a plasma concentration of sphingosine-1-phosphate that is less than the median sphingosine-1-phosphate plasma concentration in healthy volunteers. The plasma concentrations were measured in the same experimental protocol in subjects and healthy volunteers.

In one embodiment, the subject exhibits a plasma concentration of sphingosine-1-phosphate that is at least 10% or at least 15% lower than the median plasma concentration of sphingosine-1-phosphate in healthy volunteers. The plasma concentrations were measured in the same experimental protocol in subjects and healthy volunteers.

In one embodiment, the subject exhibits a plasma concentration of sphingosine-1-phosphate of less than 700nM, preferably less than 650nM, 600nM, 550nM, 500nM, 450nM, 400nM, 350nM, 300nM, 250nM, 200nM, 150nM, 100nM, 50nM or 10 nM. Preferably, the subject exhibits a plasma concentration of sphingosine-1-phosphate of less than 650 nM.

In one embodiment, the subject is a male subject and exhibits a plasma concentration of sphingosine-1-phosphate of less than 410 nM. In one embodiment, the subject is a female subject and exhibits a plasma concentration of sphingosine-1-phosphate of less than 350 nM.

In one embodiment, the subject is a male subject and exhibits a plasma concentration of sphingosine-1-phosphate that is at least 5% or at least 10% lower than 410 nM. In one embodiment, the subject is a female subject and exhibits a plasma concentration of sphingosine-1-phosphate that is at least 5% or at least 10% lower than 350 nM.

Compounds for use

The compounds used according to the invention are sphingosine-1-phosphate pathway activators selected from sphingosine-1-phosphate (S1P), sphingosine-1-phosphate lyase inhibitors, sphingosine-1-phosphate receptor agonists or sphingosine kinase activators.

In a first embodiment, the compound used is sphingosine-1-phosphate.

According to one variant, the compound used is sphingosine.

In a second embodiment, the compound used is selected from sphingosine-1-phosphate lyase inhibitors.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-1-deoxy-dihydrosphingosine-1-phosphonic acid,

-2-vinyldihydrosphingosine-1-phosphate,

-N- (4- (1H-imidazol-1-yl) benzyl) -4-amino-5-oxo-2- (piperidin-1-yl) -8- ((tetrahydrofuran-2-yl) methyl) -5, 8-dihydropyrido [2,3-d ] pyrimidine-6-carboxamide,

- (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine,

- (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile,

- (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile,

-5- (hydroxymethyl) -2, 4-dimethylpiperidin-3-ol (known as 4-deoxypyridoxine), and

-a compound of formula (I),

wherein

R₁Represents H, acetyl, propionyl, N-hydroxyaminoacetyl, isoAzol-3-yl, 4, 5-dihydroisoOxazol-3-yl or prop-1-en-2-yl; said different isAzol-3-yl or 4, 5-dihydroisoThe oxazol-3-yl is optionally substituted with at least one alkyl, acetyl, haloalkyl, hydroxyalkyl, alkoxy, alkylamino, or nitro group.

R₂Represents 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, ethyl carboxylate, N-methylformamide, 1,2-dihydroxyethyl, 1-hydroxyethyl, 2-hydroxypropan-2-yl or a phosphate thereof; and

R₃represents H, methyl, ethyl, methoxy or alkoxyalkyl.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-N- (4- (1H-imidazol-1-yl) benzyl) -4-amino-5-oxo-2- (piperidin-1-yl) -8- ((tetrahydrofuran-2-yl) methyl) -5, 8-dihydropyrido [2,3-d ] pyrimidine-6-carboxamide,

- (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine,

- (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile,

- (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile, and

-a compound of formula (I) wherein

R₁Represents acetyl, N-hydroxyaminoacetyl or isoAzol-3-yl, 4, 5-dihydroisoOxazol-3-yl or prop-1-en-2-yl;

R₂represents a 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, 1, 2-dihydroxyethyl, 1-hydroxyethyl or 2-hydroxypropan-2-yl group; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

- (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine,

- (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile,

- (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile, and

-a compound of formula (I) wherein

R₁Represents acetyl, N-hydroxyaminoacetyl or isoAzol-3-yl or 4, 5-dihydroisoOxazol-3-yl;

R₂represents a 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, 1, 2-dihydroxyethyl, 1-hydroxyethyl or 2-hydroxypropan-2-yl group; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

- (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine,

- (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile,

- (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile, and

-a compound of formula (I) wherein

R₁Represents acetyl, N-hydroxyaminoacetyl or isoOxazol-3-yl;

R₂represents a 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, 1, 2-dihydroxyethyl, 1-hydroxyethyl or 2-hydroxypropan-2-yl group; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

- (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine,

- (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile,

- (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile, and

-a compound of formula (I) wherein

R₁Represents acetyl or N-hydroxyaminoacetyl;

R₂represents a 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, 1, 2-dihydroxyethyl, 1-hydroxyethyl or 2-hydroxypropan-2-yl group; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

- (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine,

- (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile, and

- (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-a compound of formula (I),

wherein

R₁Represents acetyl, N-hydroxyaminoacetyl or isoAzol-3-yl or 4, 5-dihydroisoOxazol-3-yl;

R₂represents a 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, 1, 2-dihydroxyethyl, 1-hydroxyethyl or 2-hydroxypropan-2-yl group; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-a compound of formula (I) wherein

R₁Represents acetyl, N-hydroxyaminoacetyl or isoOxazol-3-yl;

R₂represents 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl or H; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-a compound of formula (I) wherein

R₁Represents acetyl, N-hydroxyaminoacetyl or isoOxazol-3-yl;

R₂represents 1,2,3, 4-tetrahydroxybutyl or hydroxymethyl; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-a compound of formula (I) wherein

R₁Represents acetyl or N-hydroxyaminoacetyl;

R₂represents 1,2,3, 4-tetrahydroxybutyl or hydroxymethyl; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-a compound of formula (I) wherein

R₁Represents acetyl or N-hydroxyaminoacetyl;

R₂represents a hydroxymethyl group; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of THI (2-acetyl-4 (5) -tetrahydroxybutyl-imidazole), metabolites and derivatives thereof. Thus, the sphingosine-1-phosphate lyase inhibitor may be selected from:

-a compound of formula (I) wherein

R₁Represents an acetyl group;

R₂represents a 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, 1, 2-dihydroxyethyl, 1-hydroxyethyl or 2-hydroxypropan-2-yl group; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-a compound of formula (I) wherein

R₁Represents an acetyl group;

R₂represents 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl or H; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-a compound of formula (I) wherein

R₁Represents an acetyl group;

R₂represents 1,2,3, 4-tetrahydroxybutyl or hydroxymethyl; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of L X2931(1- (4- (-1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime), metabolites and derivatives thereof.

-a compound of formula (I),

wherein

R₁Represents N-hydroxyaminoacetyl;

R₂represents a 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, 1, 2-dihydroxyethyl, 1-hydroxyethyl or 2-hydroxypropan-2-yl group; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-a compound of formula (I) wherein

R₁Represents N-hydroxyaminoacetyl;

R₂represents 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl or H; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-a compound of formula (I) wherein

R₁Represents N-A hydroxyacetamidoyl group;

R₂represents 1,2,3, 4-tetrahydroxybutyl or hydroxymethyl; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of L X2932(1- (2- (iso-phospho-lyase)Oxazol-3-yl) -1H-imidazol-4-yl) 1,2,3, 4-tetrahydroxybutyl), metabolites and derivatives thereof. Thus, the sphingosine-1-phosphate lyase inhibitor may be selected from:

-a compound of formula (I),

wherein

R₁Means for indicating differentOxazol-3-yl;

R₂represents a 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl, H, 1, 2-dihydroxyethyl, 1-hydroxyethyl or 2-hydroxypropan-2-yl group; and

R₃represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor may be selected from the group consisting of:

-a compound of formula (I) wherein

R₁Means for indicating differentOxazol-3-yl;

R₂represents 1,2,3, 4-tetrahydroxybutyl, hydroxymethyl or H; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor may be selected from the group consisting of:

-a compound of formula (I) wherein

R₁Means for indicating differentOxazol-3-yl;

R₂represents 1,2,3, 4-tetrahydroxybutyl or hydroxymethyl; and is

R₃Represents H or alkyl, preferably R₃Represents H or methyl, even more preferably R₃Represents H.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-1-deoxy-dihydrosphingosine-1-phosphonic acid,

-2-vinyldihydrosphingosine-1-phosphate,

-N- (4- (1H-imidazol-1-yl) benzyl) -4-amino-5-oxo-2- (piperidin-1-yl) -8- ((tetrahydrofuran-2-yl) methyl) -5, 8-dihydropyrido [2,3-d ] pyrimidine-6-carboxamide,

- (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine and derivatives thereof,

- (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile,

- (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile,

-5- (hydroxymethyl) -2, 4-dimethylpiperidin-3-ol (known as 4-deoxypyridoxine),

-2-acetyl-4 (5) -tetrahydroxybutyl-imidazole (known as THI),

- (E) -1- (4- ((1R,2S,3R) -1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime (known as L X2931),

- (1R,2S,3R) -1- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol (designated L X2932),

- (1R,2S,3R) -1- (2- (4, 5-dihydroiso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol,

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone,

-1- (1H-imidazol-2-yl) ethanone,

-2-acetyl-1H-imidazole-4-carboxylic acid ethyl ester,

-2-acetyl-N-methyl-1H-imidazole-4-carboxamide,

-1- (4- (1, 2-dihydroxyethyl) -1H-imidazol-2-yl) ethanone,

-1- (4- (1-hydroxyethyl) -1H-imidazol-2-yl) ethanone,

-1- (4- (2-hydroxypropan-2-yl) -1H-imidazol-2-yl) ethanone,

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone,

-1- (4- (hydroxymethyl) -1-methyl-1H-imidazol-2-yl) ethanone,

-1- (1-ethyl-4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone,

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) propan-1-one,

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone oxime,

- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) methanol, and

- (2- (prop-1-en-2-yl) -1H-imidazol-4-yl) methanol.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

- (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine,

- (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile,

- (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile,

-5- (hydroxymethyl) -2, 4-dimethylpiperidin-3-ol (known as 4-deoxypyridoxine),

-2-acetyl-4 (5) -tetrahydroxybutyl-imidazole (known as THI),

- (E) -1- (4- ((1R,2S,3R) -1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime (known as L X2931),

- (1R,2S,3R) -1- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol (designated L X2932),

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone,

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) propan-1-one,

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone oxime, and

- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) methanol.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-1-deoxy-dihydrosphingosine-1-phosphonic acid,

-2-vinyldihydrosphingosine-1-phosphate,

-N- (4- (1H-imidazol-1-yl) benzyl) -4-amino-5-oxo-2- (piperidin-1-yl) -8- ((tetrahydrofuran-2-yl) methyl) -5, 8-dihydropyrido [2,3-d ] pyrimidine-6-carboxamide,

- (R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine and derivatives thereof,

- (R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile,

- (R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile,

-5- (hydroxymethyl) -2, 4-dimethylpiperidin-3-ol (known as 4-deoxypyridoxine),

-2-acetyl-4 (5) -tetrahydroxybutyl-imidazole (known as THI),

- (E) -1- (4- ((1R,2S,3R) -1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime (known as L X2931),

- (1R,2S,3R) -1- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol (designated L X2932), and

- (1R,2S,3R) -1- (2- (4, 5-dihydroiso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-2-acetyl-4 (5) -tetrahydroxybutyl-imidazole (known as THI),

- (E) -1- (4- ((1R,2S,3R) -1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime (designated L X2931), and

- (1R,2S,3R) -1- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol,

-or a combination thereof.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is L X2931(1- (4- (1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime.) in one embodiment, the sphingosine-1-phosphate lyase inhibitor is L X2932(1- (2- (iso-hydroxybutyl) -1H-imidazol-2-yl) ethanone oxime)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol). In one embodiment, the sphingosine-1-phosphate lyase inhibitor is 2-acetyl-4 (5) -tetrahydroxybutyl-imidazole. In one embodiment, the sphingosine-1-phosphate lyase inhibitor is 1- (4- (1-hydroxymethyl) -1H-imidazol-2-yl) ketene.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of the compounds of formula (I) described previously and active metabolites thereof.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-2-acetyl-4 (5) -tetrahydroxybutyl-imidazole,

-1- (4- (1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime,

-1- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol,

-1- (4- (1-hydroxymethyl) -1H-imidazol-2-yl) ethanone,

-1- (4- (1-hydroxyethyl) -1H-imidazol-2-yl) ethanone,

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone oxime, and

- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) methanol.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-2-acetyl-4 (5) -tetrahydroxybutyl-imidazole,

-1- (4- (1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime,

-1- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol,

-1- (4- (1-hydroxyethyl) -1H-imidazol-2-yl) ethanone,

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone oxime, and

- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) methanol.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-2-acetyl-4 (5) -tetrahydroxybutyl-imidazole,

-1- (4- (1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime,

-1- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol,

-1- (4- (1-hydroxymethyl) -1H-imidazol-2-yl) ethanone,

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone oxime, and

- (2- (iso)Oxazol-3-yl) -1H-imidazol-4-yl) methanol.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-2-acetyl-4 (5) -tetrahydroxybutyl-imidazole,

-1- (4- (1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime,

-1- (4- (1-hydroxyethyl) -1H-imidazol-2-yl) ethanone, and

-1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone oxime.

In one embodiment, the sphingosine-1-phosphate lyase inhibitor is selected from the group consisting of:

-2-acetyl-4 (5) -tetrahydroxybutyl-imidazole,

-1- (4- (1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime, and

-1- (4- (1-hydroxyethyl) -1H-imidazol-2-yl) ethanone.

Use of the CompoundsUltra version 12.0(Perkinelmer) name.

N- (4- (1H-imidazol-1-yl) benzyl) -4-amino-5-oxo-2- (piperidin-1-yl) -8- ((tetrahydrofuran-2-yl) methyl) -5, 8-dihydropyrido [2,3-d ] pyrimidine-6-carboxamide

(R) -1-benzyl-4- (3-methyl-4- (p-tolyl) piperazin-1-yl) phthalazine

(R) -4- (4- (4-benzyl-7-chlorophthalazin-1-yl) -2-methylpiperazin-1-yl) benzonitrile

(R) -1-benzyl-4- (4- (4-cyanophenyl) -3-methylpiperazin-1-yl) phthalazine-6-carbonitrile

5- (hydroxymethyl) -2, 4-dimethylpiperidin-3-ol

2-acetyl-4 (5) -tetrahydroxybutyl-imidazole (known as THI)

(E) -1- (4- ((1R,2S,3R) -1,2,3, 4-tetrahydroxybutyl) -1H-imidazol-2-yl) ethanone oxime (named L X2931)

(1R,2S,3R) -1- (2- (4, 5-dihydroiso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol

(1R,2S,3R) -1- (2- (iso-iso)Oxazol-3-yl) -1H-imidazol-4-yl) butane-1, 2,3, 4-tetraol

1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone

1- (4- (1, 2-dihydroxyethyl) -1H-imidazol-2-yl) ethanone

1- (4- (hydroxymethyl) -1H-imidazol-2-yl) ethanone oximes

(2- (iso-iso)Oxazol-3-yl) -1H-imidazol-4-yl) methanol

In one embodiment, the sphingosine kinase activator is calcitriol (1 α, 25-dihydroxy vitamin D)₃)。

In a fourth embodiment, the compound used is selected from sphingosine-1-phosphate receptor agonists. In one embodiment, the sphingosine-1-phosphate receptor agonist is selected from the group consisting of:

-2-amino-2- (4-octylphenethyl) propane-1, 3-diol (known as Fingolimod or FTY720),

-4- (4- (5- (5-chloro-6-isopropoxypiperidin-3-yl) -1,2,4-Oxadiazol-3-yl) -1H-indol-1-yl) butanoic acid(referred to as GSK 2018682) is,

- (2Z,5Z) -5- (3-chloro-4- ((R) -2, 3-dihydroxypropoxy) benzylidene) -2- (propylimino) -3- (o-tolyl) thiazolidin-4-one (known as Ponesimod),

-KRP-203，

-KRP-203，

-AUY954，

-ML178，

-XAX162，

-ML248，

-SEW2871，

-CYM5442，

-CYM5441，

-CYM5520，

-RP001，

-ASP4058，

-CS-0777，

-CS2100，

-BAF312，

-SID 46371153，

-AMG369, and

-phosphorylated derivatives thereof.

In one embodiment, the sphingosine-1-phosphate receptor agonist is selected from the group consisting of:

-2-amino-2- (4-octylphenethyl) propane-1, 3-diol (known as Fingolimod or FTY720),

-4- (4- (5- (5-chloro-6-isopropoxypiperidin-3-yl) -1,2,4-Oxadiazol-3-yl) -1H-indol-1-yl) butanoic acid (known as GSK 2018682),

- (2Z,5Z) -5- (3-chloro-4- ((R) -2, 3-dihydroxypropoxy) benzylidene) -2- (propylimino) -3- (o-tolyl) thiazolidin-4-one (known as Ponesimod), and

-phosphorylated derivatives thereof.

In one embodiment, the sphingosine-1-phosphate receptor agonist is selected from FTY720 and phosphorylated FTY720((S) - (2-amino-2- (hydroxymethyl) -4- (4-octylphenyl) butyl) phosphonic acid or FTY 720P). In one embodiment, the sphingosine-1-phosphate receptor agonist is FTY 720. In one embodiment, the sphingosine-1-phosphate receptor agonist is FTY 720P. In one embodiment, the sphingosine-1-phosphate receptor agonist is GSK 2018682. In one embodiment, the sphingosine-1-phosphate receptor agonist is Ponesidmod.

Use of the CompoundsUltra version 12.0(Perkinelmer) name.

2-amino-2- (4-octylphenethyl) propane-1, 3-diol (known as FTY720 or Fingolimod)

(S) - (2-amino-2- (hydroxymethyl) -4- (4-octylphenyl) butyl) phosphonic acid (named FTY20P)

4- (4- (5- (5-chloro-6-isopropoxypiperidin-3-yl) -1,2,4-Oxadiazol-3-yl) -1H-indol-1-yl butyric acid (known as GSK 2018682)

(2Z,5Z) -5- (3-chloro-4- ((R) -2, 3-dihydroxypropoxy) benzylidene) -2- (propylimino) -3- (o-tolyl) thiazolidin-4-one (named Ponesimod)

Method of treatment

The present invention relates to compounds useful as neoadjuvants in methods of treating cancer in a subject.

The subject is a mammal, preferably a human. In one embodiment, the subject may be waiting for or receiving medical services, or have/is/will be targeted for medical procedures, or be monitored for disease development.

In one embodiment, the subject is an adult (e.g., a subject older than 18 years of age). In another embodiment, the subject is a child (e.g., a subject younger than 18 years of age). In one embodiment, the subject is male. In another embodiment, the subject is female.

In one embodiment, a subject is first treated according to the methods of the present invention. In one embodiment, the subject is not responsive to the first chemotherapy or radiation therapy.

The compounds used as neoadjuvant are administered prior to the sequential chemotherapy. In one embodiment, the compound used as neoadjuvant is administered at least once prior to the sequential chemotherapy. In another embodiment, the neoadjuvant of the present invention is administered in repeated doses prior to sequential chemotherapy.

In one embodiment, at least one administration of a compound used as neoadjuvant of the present invention is performed 16 weeks, 15 weeks, 14 weeks, 13 weeks, 12 weeks, 11 weeks, 10 weeks, 9 weeks, 8 weeks, 7 weeks, 6 weeks, 5 weeks, 4 weeks, 3 weeks, 2 weeks or 1 week prior to the sequential chemotherapy. In one embodiment, at least one administration of a compound used as neoadjuvant of the present invention is performed 7 days, 6 days, 5 days, 4 days, 3 days, 2 days or 1 day before the sequential chemotherapy. In one embodiment, at least one administration of a compound used as a neoadjuvant of the present invention is performed 24 hours, 23 hours, 22 hours, 21 hours, 20 hours, 19 hours, 18 hours, 17 hours, 16 hours, 15 hours, 14 hours, 13 hours, 12 hours, 11 hours, 10 hours, 9 hours, 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, or 1 hour prior to the sequential chemotherapy.

In one embodiment, the neoadjuvant is administered as a pharmaceutical composition comprising at least one compound for use according to the invention and one or more than one pharmaceutically acceptable diluent or carrier for the subject.

The novel adjuvants may be administered by any conventional route, in particular enterally, for example orally, for example in the form of tablets, capsules, drink solutions, intranasally, pulmonarily or parenterally, for example in the form of injection solutions or suspensions.

Suitable unit dosage forms for oral administration comprise 0.01mg to 1000mg together with one or more pharmaceutically acceptable diluents or carriers. To inhibit sprouting and stabilize neovascular connections in tumors, an appropriate dose to provide vascular stabilization when the neoadjuvant is administered to a subject can be selected by concentration and dose escalation studies as described in the art. In one embodiment, the dose of neoadjuvant is adjusted to achieve a plasma concentration of sphingosine-1-phosphate greater than 650 nM.

According to the present invention, a method of treating cancer comprises administering sequential chemotherapy to a subject after administration of a neoadjuvant. In one embodiment, the administration of the sequential chemotherapy comprises administration of at least one anti-cancer agent. In one embodiment, the at least one anticancer drug is selected from paclitaxel, gemcitabine, and oxaliplatin.

According to another embodiment, the anti-cancer agent is a single agent or a combination of anti-cancer agents, provided that none of these agents exhibit anti-angiogenic properties. According to this embodiment, the anti-cancer agent is not a VEGF inhibitor, nor an EGFR inhibitor, nor a HER2 inhibitor, nor bevacizumab, nor cetuximab, nor panitumumab, nor trastuzumab. According to this embodiment, the anti-cancer agent is not a VEGFR-1 inhibitor, nor a VEGFR-2 inhibitor, nor a VEGFR-3 inhibitor, nor a PDGF inhibitor, nor a c-kit inhibitor, nor a STAT3 inhibitor, nor a FGFR inhibitor, nor a RAF inhibitor, nor a TIMP 1 inhibitor, nor a TIMP 2 inhibitor, nor a TIMP 3 inhibitor, nor a metalloproteinase inhibitor, nor a cilansil, nor erlotinib, nor sorafenib, nor sunitinib, nor gefitinib, nor lapatinib, nor pazopanib, nor masitinib, nor imatinib.

In a second aspect, the present invention relates to a method of treating cancer using sphingosine-1-phosphate (S1P) pathway activators as neoadjuvant prior to anti-cancer treatment to improve the efficacy of anti-cancer drugs as described above.

According to a first embodiment, the S1P pathway activator inhibits sprouting angiogenesis and stabilizes the blood vessels of the tumor.

According to a second embodiment, the sphingosine-1-phosphate pathway activator increases the plasma concentration of sphingosine-1-phosphate.

According to a third embodiment, the sphingosine-1-phosphate pathway activator enhances the therapeutic effect of the anti-cancer drug.

In a third aspect, the present invention relates to a method of improving the efficacy of treating a cancerous tumor in a subject in need thereof, wherein:

a. in a first step, measuring the plasma sphingosine-1-phosphate concentration of the subject,

b. in a second step, an anti-cancer agent is administered to a subject with high S1P plasma levels,

provided that the subject does not receive an anti-angiogenic drug.

In one embodiment, the treatment is chemotherapy treatment. In one embodiment, the treatment is a radiation therapy treatment.

The plasma concentration value of sphingosine-1-phosphate can be determined according to the analytical method and sex.

In one embodiment, the subject exhibits a plasma concentration of sphingosine-1-phosphate that is less than the median plasma concentration of sphingosine-1-phosphate in healthy volunteers. The plasma concentrations were measured in the same experimental protocol in subjects and healthy volunteers.

In one embodiment, the subject exhibits a plasma concentration of sphingosine-1-phosphate that is at least 10% or at least 15% lower than the median plasma concentration of sphingosine-1-phosphate in healthy volunteers. The plasma concentrations were measured in the same experimental protocol in subjects and healthy volunteers.

In one embodiment, as previously described, if the plasma concentration of sphingosine-1-phosphate in a subject is less than 650nM after the first step, the subject receives a neoadjuvant administration prior to the second step.

The method of treatment according to any aspect of the invention may also be applied in combination with further surgical intervention, mild prolonged whole body hyperthermia and/or radiotherapy.

The method of treatment according to any aspect of the invention produces beneficial effects such as slowing, arresting or reversing neoplasm formation, metastatic spread or growth, or prolonging the duration of tumor response. It may also lead to other beneficial effects, such as limiting side effects, improving quality of life or reducing mortality and morbidity, compared to monotherapy using only chemotherapeutic drugs.

The method of treatment according to any aspect of the invention relates to any type of cancer. In a preferred embodiment, the cancer is a solid tumor cancer. In one embodiment, the cancer is selected from tumors including brain and other central nervous system, head cancer and/or neck cancer; breast tumors; tumors of the circulatory system (e.g., heart, mediastinum, and pleura, as well as other organs within the thoracic cavity, vascular tumors, and vascular tissue associated with tumors); tumors of the excretory system (e.g., kidney, renal pelvis, ureter, bladder, other unspecified urinary organs); gastrointestinal tumors (e.g., esophagus, stomach, small intestine, colon, colorectal, rectosigmoid junction, rectum, anus, and anal canal), tumors involving the liver and intrahepatic bile ducts, gallbladder, other unspecified parts of the biliary tract, pancreas, other digestive organs); oral cavity (lips, tongue, gums, floor of mouth, palate and other parts of oral cavity, parotid gland, other parts of salivary glands, tonsil, oropharynx, nasopharynx, pyriform sinus, hypopharynx and other parts of lips, oral cavity and pharynx); tumors of the reproductive system; tumors of the respiratory tract (e.g., nasal and middle ear, paranasal sinuses, larynx, trachea, bronchi, and lung, e.g., small cell lung cancer or non-small cell lung cancer); skeletal system tumors (e.g., bone and articular cartilage of the extremities, osteoarticular cartilage, and other sites); skin tumors (e.g., malignant melanoma of the skin, non-melanoma skin cancer, basal cell carcinoma of the skin, squamous cell carcinoma of the skin, mesothelioma, kaposi's sarcoma); and tumors involving other tissues including the peripheral and autonomic nervous systems, connective and soft tissues, the retroperitoneal cavity and peritoneum, the eye and appendages, the thyroid, adrenal glands and other endocrine glands and related structures, secondary and unspecified malignancies of the lymph nodes, secondary malignancies of the respiratory and digestive systems, and secondary malignancies in other locations. Where a tumor, tumor disease, cancer, or cancer is mentioned above and subsequently, metastasis of the original organ or tissue and/or any other location may alternatively or additionally be implied, regardless of the location of the tumor and/or metastasis. In a preferred embodiment, the treatment according to any aspect of the invention relates to the treatment of pancreatic cancer, preferably pancreatic cancer.

Prognosis of solid tumor treatment outcome

In one aspect, the invention relates to sphingosine-1-phosphate for use in an in vitro non-invasive method for predicting the clinical outcome of treatment of a subject. Wherein the method comprises the following steps:

a) detecting and quantifying the concentration of sphingosine-1-phosphate in a plasma sample previously obtained from a subject, an

b) Comparing the results obtained in step a) with the median plasma sphingosine-1-phosphate concentration in healthy volunteers; the plasma concentrations were measured in the same experimental protocol in subjects and healthy volunteers.

In one embodiment, the method comprises:

a) detecting and quantifying the concentration of sphingosine-1-phosphate in a plasma sample previously obtained from a subject, an

b) Comparing the result obtained in step a) with a reference value. The reference value is at least 10% or at least 15% lower than the median sphingosine-1-phosphate plasma concentration in healthy volunteers; the plasma concentrations were measured in the same experimental protocol in subjects and healthy volunteers.

In one embodiment, the method comprises:

a) detecting and quantifying the concentration of sphingosine-1-phosphate in a plasma sample previously obtained from a subject, an

b) Comparing the result obtained in step a) with a reference value of 650nM for sphingosine-1-phosphate.

In one embodiment, the method comprises:

a) detecting and quantifying the concentration of sphingosine-1-phosphate in a plasma sample previously obtained from a subject, an

b) Comparing the result obtained in step a) with a reference value for sphingosine-1-phosphate, which is 410nM if the subject is male and 350nM if the subject is female.

In one embodiment, the treatment is chemotherapy treatment. In one embodiment, the treatment is a radiation therapy treatment.

In one embodiment, the subject has pancreatic cancer.

In another aspect, the invention relates to an in vitro non-invasive method for predicting the clinical outcome of a treatment of a subject. Wherein the method comprises the following steps:

a) detecting and quantifying the concentration of sphingosine-1-phosphate in a plasma sample previously obtained from a subject, an

b) Comparing the result obtained in step a) with a reference value of 650nM for sphingosine-1-phosphate.

In one embodiment, the treatment is chemotherapy. In one embodiment, as previously described, the chemotherapy does not include drugs with anti-angiogenic properties.

In one embodiment, the subject is to receive chemotherapy. In another embodiment, the subject is undergoing chemotherapy during the in vitro method of the invention.

Applicants have shown that subjects exhibiting plasma concentrations of sphingosine-1-phosphate above 650nM show better clinical results after administration of chemotherapy.

Solid tumor prognosis kit

In another aspect, the invention relates to a solid tumor prognosis kit for performing the in vitro non-invasive method of the invention, consisting of a set of reagents suitable for purifying and/or quantifying sphingosine-1-phosphate.

In one embodiment, the reagent is selected from the group consisting of solvents, buffers, standard solutions of sphingosine-1-phosphate, antibodies specific for sphingosine-1-phosphate, labeled antibodies specific for sphingosine-1-phosphate, sphingosine-1-phosphate coated E. L. I.S.A (enzyme linked immunosorbent assay) plaques purification and/or quantification of sphingosine-1-phosphate can be performed using techniques generally known in the art, including but not limited to centrifugation, solvent extraction, solid phase extraction, filtration, E. L. I.S.A. and other immunoenzymatic or immunochemical techniques as well as chromatographic techniques, preferably mass spectrometry-chromatography techniques.