WO2012157746A1 - Therapeutic drug for autoimmune disease - Google Patents
Therapeutic drug for autoimmune disease Download PDFInfo
- Publication number
- WO2012157746A1 WO2012157746A1 PCT/JP2012/062782 JP2012062782W WO2012157746A1 WO 2012157746 A1 WO2012157746 A1 WO 2012157746A1 JP 2012062782 W JP2012062782 W JP 2012062782W WO 2012157746 A1 WO2012157746 A1 WO 2012157746A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- alkyl
- amino
- hydroxyphosphoryloxy
- alkoxy
- phenyl
- Prior art date
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- 208000023275 Autoimmune disease Diseases 0.000 title claims abstract description 37
- 229940126585 therapeutic drug Drugs 0.000 title 1
- 150000001875 compounds Chemical class 0.000 claims abstract description 145
- 238000012360 testing method Methods 0.000 claims abstract description 57
- 238000000034 method Methods 0.000 claims abstract description 54
- 101001040717 Homo sapiens Probable G-protein coupled receptor 174 Proteins 0.000 claims abstract description 35
- 101710085183 Putative P2Y purinoceptor 10 Proteins 0.000 claims abstract description 35
- 102100021199 Probable G-protein coupled receptor 174 Human genes 0.000 claims abstract description 29
- 230000000694 effects Effects 0.000 claims abstract description 27
- 238000012216 screening Methods 0.000 claims abstract description 24
- 239000000556 agonist Substances 0.000 claims abstract description 15
- 102000005962 receptors Human genes 0.000 claims abstract description 15
- 108020003175 receptors Proteins 0.000 claims abstract description 15
- 239000003814 drug Substances 0.000 claims abstract description 14
- 229940124597 therapeutic agent Drugs 0.000 claims abstract description 10
- 230000003449 preventive effect Effects 0.000 claims abstract description 7
- 125000000217 alkyl group Chemical group 0.000 claims description 161
- 125000003545 alkoxy group Chemical group 0.000 claims description 80
- -1 dioxolenoneylmethyl Chemical group 0.000 claims description 55
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 55
- 210000004027 cell Anatomy 0.000 claims description 33
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 31
- 150000003839 salts Chemical class 0.000 claims description 30
- 125000006570 (C5-C6) heteroaryl group Chemical group 0.000 claims description 29
- 239000003795 chemical substances by application Substances 0.000 claims description 25
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 claims description 22
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 21
- 125000004432 carbon atom Chemical group C* 0.000 claims description 21
- 125000005196 alkyl carbonyloxy group Chemical group 0.000 claims description 18
- 230000000069 prophylactic effect Effects 0.000 claims description 17
- 125000001072 heteroaryl group Chemical group 0.000 claims description 16
- 108090000623 proteins and genes Proteins 0.000 claims description 16
- 230000001225 therapeutic effect Effects 0.000 claims description 15
- 201000010099 disease Diseases 0.000 claims description 14
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 14
- 125000005549 heteroarylene group Chemical group 0.000 claims description 14
- 125000000843 phenylene group Chemical group C1(=C(C=CC=C1)*)* 0.000 claims description 14
- ZPDQFUYPBVXUKS-YADHBBJMSA-N 1-stearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@@H](O)COP(O)(=O)OC[C@H](N)C(O)=O ZPDQFUYPBVXUKS-YADHBBJMSA-N 0.000 claims description 13
- 125000005843 halogen group Chemical group 0.000 claims description 13
- 210000004698 lymphocyte Anatomy 0.000 claims description 13
- 125000005633 phthalidyl group Chemical group 0.000 claims description 13
- 125000001424 substituent group Chemical group 0.000 claims description 12
- 102000006747 Transforming Growth Factor alpha Human genes 0.000 claims description 11
- 101800004564 Transforming growth factor alpha Proteins 0.000 claims description 11
- 125000000051 benzyloxy group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])O* 0.000 claims description 11
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 11
- 235000019260 propionic acid Nutrition 0.000 claims description 11
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 claims description 11
- 102000002260 Alkaline Phosphatase Human genes 0.000 claims description 10
- 108020004774 Alkaline Phosphatase Proteins 0.000 claims description 10
- 239000003112 inhibitor Substances 0.000 claims description 10
- 230000004073 interleukin-2 production Effects 0.000 claims description 10
- 201000000596 systemic lupus erythematosus Diseases 0.000 claims description 9
- 125000004429 atom Chemical group 0.000 claims description 8
- 125000004454 (C1-C6) alkoxycarbonyl group Chemical group 0.000 claims description 7
- 125000004916 (C1-C6) alkylcarbonyl group Chemical group 0.000 claims description 7
- 125000005194 alkoxycarbonyloxy group Chemical group 0.000 claims description 7
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 claims description 6
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 claims description 6
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 claims description 6
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 6
- 208000003343 Antiphospholipid Syndrome Diseases 0.000 claims description 5
- 208000023328 Basedow disease Diseases 0.000 claims description 5
- 208000008439 Biliary Liver Cirrhosis Diseases 0.000 claims description 5
- 208000033222 Biliary cirrhosis primary Diseases 0.000 claims description 5
- 208000015023 Graves' disease Diseases 0.000 claims description 5
- 208000012654 Primary biliary cholangitis Diseases 0.000 claims description 5
- 208000021386 Sjogren Syndrome Diseases 0.000 claims description 5
- 201000006417 multiple sclerosis Diseases 0.000 claims description 5
- 206010028417 myasthenia gravis Diseases 0.000 claims description 5
- 208000005987 polymyositis Diseases 0.000 claims description 5
- 206010048628 rheumatoid vasculitis Diseases 0.000 claims description 5
- PEIJVSZKYNNUKV-MOPGFXCFSA-N (2S)-2-amino-3-[[(2R)-3-[3-(2-heptoxyphenyl)propanoyloxy]-2-hydroxypropoxy]-hydroxyphosphoryl]oxypropanoic acid Chemical compound CCCCCCCOc1ccccc1CCC(=O)OC[C@@H](O)COP(O)(=O)OC[C@H](N)C(O)=O PEIJVSZKYNNUKV-MOPGFXCFSA-N 0.000 claims description 4
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- TUOGAVKNETYDTH-IKZNXBAASA-N (2S,3S)-2-amino-3-[hydroxy-[(2R)-2-hydroxy-3-[(Z)-octadec-9-enoyl]oxypropoxy]phosphoryl]oxybutanoic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](O)COP(O)(=O)O[C@@H](C)[C@H](N)C(O)=O TUOGAVKNETYDTH-IKZNXBAASA-N 0.000 claims description 4
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- SIUKGQNAMYOCLM-FQEVSTJZSA-N (2S)-2-amino-3-[3-hexadecanoyloxypropoxy(hydroxy)phosphoryl]oxypropanoic acid Chemical compound CCCCCCCCCCCCCCCC(=O)OCCCOP(O)(=O)OC[C@H](N)C(O)=O SIUKGQNAMYOCLM-FQEVSTJZSA-N 0.000 claims description 3
- SQIXEOUVMZQPKG-NRFANRHFSA-N (2S)-2-amino-3-[3-hexadecoxypropoxy(hydroxy)phosphoryl]oxypropanoic acid Chemical compound CCCCCCCCCCCCCCCCOCCCOP(=O)(O)OC[C@@H](C(=O)O)N SQIXEOUVMZQPKG-NRFANRHFSA-N 0.000 claims description 3
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Images
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/564—Immunoassay; Biospecific binding assay; Materials therefor for pre-existing immune complex or autoimmune disease, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
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- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
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- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
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Definitions
- the present invention relates to a screening method for compounds useful as autoimmune therapeutic agents.
- the present invention also relates to a therapeutic agent for autoimmune diseases.
- the invention further relates to lysophosphatidylserine and novel derivatives thereof.
- Lysophospholipid is a general term for phospholipids having one acyl group.
- the lysophospholipid has a property that it is less hydrophobic than the diacylphospholipid constituting the cell membrane and can be easily released from the cell membrane.
- Some lysophospholipids function as signal molecules between cells or between membranes and have been found to have an important role in vivo.
- Patent Document 1 Japanese Patent Application Laid-Open No. 2007-267601.
- lysophosphatidylserine which is a kind of lysophospholipid, is involved in acute inflammation caused by mast cell degranulation (Non-patent Documents 1 and 2).
- LPS 1 GPR34
- LPS 2 P2Y10
- LPS 3 A630033H20Rik
- LPS 4 GPR174
- LPS 1 is involved in signal transduction that induces or enhances degranulation of mast cells and can be a target for the treatment of allergic diseases and chronic inflammatory diseases
- Patent Document 2 a specific lysophosphatidylserine derivative
- lysophosphatidylthreonine strongly promotes the degranulation reaction of mast cells
- Autoimmune disease is a general term for diseases that cause symptoms when the immune system, which is originally a defense mechanism against foreign substances, reacts excessively and attacks even normal cells and tissues of itself.
- systemic autoimmune diseases that affect the whole body and organ-specific diseases that affect only specific organs.
- autoimmune diseases often become chronic diseases or intractable diseases, and some of them are designated as diseases for which specific diseases are researched by the Ministry of Health, Labor and Welfare.
- Much research has been conducted on methods for treating autoimmune diseases, for example, methods for treating chronic inflammation caused by autoimmune diseases using cytokine-specific antibodies involved in inflammation (Patent Document 2), and pathogenic self.
- Patent Document 3 A method for treating diseases by neutralizing antibodies
- autoimmune diseases many causes of autoimmune diseases have not been elucidated, and effective treatments have not yet been established for many autoimmune diseases.
- drugs that suppress the immune system and anti-inflammatory drugs stereos or non-steroid drugs that relieve inflammation are used as the first selection agent.
- An object of the present invention is to provide a new treatment method for autoimmune diseases. Furthermore, the present invention is to provide a method for assaying compounds useful for the treatment of autoimmune diseases.
- the inventors of the present invention have made extensive studies in order to achieve the above-mentioned problems. As a result, the compounds having agonist activity of LPS 2 (P2Y10) and LPS 4 (GPR174) have been found to have therapeutic and preventive effects on autoimmune diseases. Completed the invention.
- the following screening method is provided.
- a screening method for a therapeutic or prophylactic agent for an autoimmune disease comprising using P2Y10 or GPR174 to evaluate the agonist activity or antagonist activity of a test compound for a lysophosphatidylserine receptor.
- the method according to (1) above wherein the agonist activity is evaluated.
- the method according to (1) or (2) above for screening for an interleukin 2 production inhibitor.
- EGFR human epidermal growth factor receptor
- AP alkaline phosphatase
- TGF ⁇ transforming growth factor ⁇
- R 11 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, or benzyl
- R 12 and R 13 are each independently a hydrogen atom, C 1-6 alkyl, formyl, C 1-6 alkylcarbonyl, C 1-6 alkoxycarbonyl, C 1-6 alkoxy C 1-6 alkyl, or benzyloxy Selected from carbonyl
- R 15 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, C 1-6 alkylcarbonyloxy C 1-6 alkyl, C 1-6 alkoxycarbonyloxy C 1-6 alkyl, Phenyl, benzyl, phthalidyl, dioxolenoneylmethyl, or furylmethyl
- p is an integer selected from 2 to 6
- Y 1 is a direct bond or —C ( ⁇ O) —
- R 16 is C 3-30 alkyl, wherein one or more phen
- the phenyl or heteroaryl may be substituted with one or more substituents selected from C 1-10 alkyl, C 1-10 alkoxy and a halogen atom;
- the above benzyl, benzyloxy, phenyl, phthalidyl, dioxolenoneylmethyl, furylmethyl, phenylene, and heteroarylene are C 1-6 alkyl, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro May be substituted with one or more substituents selected from phenyl and halogen atoms] Or a pharmaceutically acceptable salt thereof.
- R 16 is C 10-24 alkyl (wherein one or more —CH 2 — of alkyl may be replaced by —O—, and one or more single bonds between carbon atoms are double bonds) Or may be replaced by a triple bond), or selected from the formula: — (CH 2 ) r —Q 1 ; r is an integer selected from 2 to 20; Q 1 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-10 alkyl, phenyl C 1-10 alkoxy, 5- The compound according to the above (11), or a pharmaceutically acceptable salt thereof, which may be substituted with 6-membered heteroaryl C 1-10 alkyl or 5-6-membered heteroaryl C 1-10 alkoxy.
- R 16 is C 12-20 alkyl (wherein alkyl 1 to 3 —CH 2 — may be replaced by —O—, and one single bond between carbon atoms is a double bond) Or is selected from the formula: — (CH 2 ) r —Q 1 ; r is an integer selected from 2 to 7; Q 1 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5- The compound according to the above (12), or a pharmaceutically acceptable salt thereof, which may be substituted by 6-membered heteroaryl C 1-3 alkyl or 5-6-membered heteroaryl C 1-3 alkoxy.
- R 21 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, or benzyl
- R 22 and R 23 are each independently a hydrogen atom, C 1-6 alkyl, formyl, C 1-6 alkylcarbonyl, C 1-6 alkoxycarbonyl, C 1-6 alkoxy C 1-6 alkyl, or benzyloxy Selected from carbonyl
- R 24 is a hydrogen atom or C 1-3 alkyl
- R 25 represents a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, C 1-6 alkylcarbonyloxy C 1-6 alkyl, C 1-6 alkoxycarbonyloxy C 1-6 alkyl, Phenyl, benzyl, phthalidyl, dioxolenoneylmethyl, or furylmethyl
- X 2 is a hydrogen atom, a halogen atom, hydroxy, C 1-6 alkoxy, formyloxy
- a 2 is the following formula:
- R 21 , R 22 and R 23 are as defined in (14) above, or a pharmaceutically acceptable salt thereof.
- R 26 is C 12-20 alkyl (wherein 1 to 3 —CH 2 — of alkyl may be replaced by —O—, and one single bond between carbon atoms is a double bond) Or is selected from the formula: — (CH 2 ) s —Q 2 ; s is an integer selected from 2 to 7; Q 2 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5- The compound according to the above (14) or (15), or a pharmaceutically acceptable salt thereof, which may be substituted with 6-membered heteroaryl C 1-3 alkyl, or 5-6-membered heteroaryl C 1-3 alkoxy .
- lysophosphatidylserine derivatives have therapeutic and preventive effects on autoimmune diseases, and have completed the present invention. That is, according to still another aspect of the present invention, the following therapeutic or preventive agent is provided.
- R 1 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, or benzyl
- R 2 and R 3 are each independently a hydrogen atom, C 1-6 alkyl, formyl, C 1-6 alkylcarbonyl, C 1-6 alkoxycarbonyl, C 1-6 alkoxy C 1-6 alkyl, or benzyloxy Selected from carbonyl
- R 4 is a hydrogen atom or C 1-3 alkyl
- R 5 represents a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, C 1-6 alkylcarbonyloxy C 1-6 alkyl, C 1-6 alkoxycarbonyloxy C 1-6 alkyl, Phenyl, benzyl, phthalidyl, dioxolenoneylmethyl, or furylmethyl
- X is a hydrogen atom, a halogen atom, hydroxy, C 1-6 alkoxy, formyloxy,
- the phenyl or heteroaryl may be substituted with one or more substituents selected from C 1-10 alkyl, C 1-10 alkoxy and a halogen atom;
- R 7 is a hydrogen atom or C 1-6 alkyl;
- the above benzyl, benzyloxy, phenyl, phthalidyl, dioxolenoneylmethyl, furylmethyl, phenylene, and heteroarylene are C 1-6 alkyl, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro May be substituted with one or more substituents selected from phenyl and halogen atoms]
- the therapeutic agent or preventive agent of an autoimmune disease containing the compound represented by these, or its pharmaceutically acceptable salt.
- R 6 is C 10-24 alkyl (wherein one or more —CH 2 — of alkyl may be replaced by —O—, and one or more single bonds between carbon atoms are double bonds) Or may be replaced by a triple bond), or selected from the formula: — (CH 2 ) m —Q; m is an integer selected from 2 to 20; Q is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-10 alkyl, phenyl C 1-10 alkoxy, 5-6
- R 6 is C 12-20 alkyl (wherein the alkyl 1-3 —CH 2 — may be replaced by —O—, and one single bond between carbon atoms is a double bond) Or may be selected from the formula: — (CH 2 ) m —Q; m is an integer selected from 2 to 7; Q is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5-6
- the therapeutic or prophylactic agent according to the above (19) which may be substituted with a member heteroaryl C 1-3 alkyl or a 5-6 membered heteroaryl C 1-3 alkoxy.
- (21) (2S) -2-Amino-3-[(3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid; (2S) -2-Amino-3-[(3-basenoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid; (2S) -2-amino-3-[(3-Elideyloxypropoxy) -hydroxyphosphoryloxy] propionic acid; (2S) -2-amino-3-[(3-palmitoleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid; (2S) -2-amino-3-[(3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid; (2S) -2-amino-3-[(3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid; (2S) -2-amino-3-[(3-myristoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
- the autoimmune diseases are malignant rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, Graves' disease, antiphospholipid antibody syndrome, Sjogren's syndrome, primary biliary cirrhosis, polymyositis, and autoimmunity
- the compound according to any one of (11) to (16) above, or the compound of the formula (I) defined in any one of (17) to (21) above, Or the pharmaceutical composition for treating or preventing an autoimmune disease containing the pharmaceutically acceptable salt is provided.
- an interleukin 2 production inhibitor containing a pharmaceutically acceptable salt thereof is provided.
- the compound according to any one of (11) to (16) above, or the compound of the formula (I) defined in any one of (17) to (21) above Alternatively, a lymphocyte adhesion inhibitor containing a pharmaceutically acceptable salt thereof is provided.
- the compound according to any one of (11) to (16) above, or the compound of the formula (I) defined in any one of (17) to (21) above Alternatively, a method of treating or preventing autoimmune disease comprising administering to a patient a pharmaceutically acceptable salt thereof.
- a therapeutic or prophylactic agent for autoimmune diseases which is a new method for treating autoimmune diseases. Furthermore, the present invention provides an efficient screening method for compounds useful for the treatment of autoimmune diseases.
- FIG. 1 is a graph showing the evaluation test results (Test Example 1) of agonist activity of lysophosphatidylserine or its analogs against the P2Y10 receptor.
- FIG. 2 is a graph showing the evaluation test results (Test Example 1) of agonist activity of lysophosphatidylserine or its analogs against the GPR174 receptor.
- FIG. 3 is a graph showing the white blood cell count of a 17-week-old systemic lupus erythematosus spontaneously developing mouse in Test Example 2.
- FIG. 4 is a graph showing autoantibody production in 17-week-old systemic lupus erythematosus spontaneously developing mice in Test Example 2.
- FIG. 1 is a graph showing the evaluation test results (Test Example 1) of agonist activity of lysophosphatidylserine or its analogs against the GPR174 receptor.
- FIG. 3 is a graph showing the white blood cell count of a 17-week-old systemic lupus
- FIG. 5 is a graph showing changes in the spleen of 17-week-old spontaneously lupus erythematosus spontaneously developing mice in Test Example 2.
- 6 is a graph showing changes in mesenteric lymph nodes in 17-week-old spontaneously lupus erythematosus mice in Test Example 2.
- FIG. 7 is a diagram showing the deposition of immune complexes in the kidneys of 17-week-old systemic lupus erythematosus spontaneously developing mice in the PBS administration group (control group) in Test Example 2.
- FIG. 8 is a graph showing the deposition of immune complexes on the kidneys of 17-week-old spontaneously lupus erythematosus spontaneously developing mice in the test compound administration group in Test Example 2.
- FIG. 9 is a graph showing blood alanine transaminase (ALT) values of human autoimmune disease hepatitis model mice in Test Example 3.
- FIG. 10 is a photograph showing the site of liver injury in the liver section of a human autoimmune disease hepatitis model mouse (PBS administration group) in Test Example 3.
- FIG. 11 is a photograph showing the site of liver injury in a liver section of a human autoimmune disease hepatitis model mouse (LPS (18: 1) administration group) in Test Example 3.
- FIG. 12 is a graph obtained by quantifying the sites of liver injury in liver sections of human autoimmune disease model mice (PBS administration group, LPS (18: 1) administration group) in Test Example 3.
- FIG. 10 is a photograph showing the site of liver injury in the liver section of a human autoimmune disease hepatitis model mouse (PBS administration group) in Test Example 3.
- FIG. 11 is a photograph showing the site of liver injury in a liver section of a human autoimmune disease hepatitis model mouse (LP
- FIG. 13 is a photograph showing the site of liver injury in a liver section of a human autoimmune disease hepatitis model mouse (deoxy-LPSLP (18: 1) administration group) in Test Example 3.
- FIG. 14 is a photograph showing the site of liver injury in a liver section of a human autoimmune disease hepatitis model mouse (LPalloTlo (18: 1) administration group) in Test Example 3.
- FIG. 15 is a graph showing the expression level of P2Y10 in spleen-derived lymphocytes activated with concanavalin A (Test Example 4).
- FIG. 16 is a graph showing the expression level of GPR174 in spleen-derived lymphocytes activated with concanavalin A (Test Example 4).
- FIG. 17 is a graph showing the influence of a test compound on adhesion of spleen-derived lymphocytes activated with concanavalin A to ICAM-1 (Test Example 5).
- FIG. 18 is a graph showing the influence of a test compound on IL-2 production of spleen-derived lymphocytes activated with concanavalin A (Test Example 6).
- FIG. 19 shows human, mouse and rat oligonucleotides used as forward and reverse primers for the P2Y10 gene (SEQ ID NO: 1-6) and oligonucleotides used as forward primer and reverse primer for the GPR174 gene (SEQ ID NO: 7- It is a figure showing the base sequence of 12).
- FIG. 20 shows the base sequence of the human P2Y10 gene (SEQ ID NO: 13).
- FIG. 21 shows the base sequence of the mouse P2Y10 gene (SEQ ID NO: 14).
- FIG. 22 is a diagram showing the base sequence of the rat P2Y10 gene (SEQ ID NO: 15).
- FIG. 23 shows the base sequence of the human GPR174 gene (SEQ ID NO: 16).
- FIG. 24 is a diagram showing the base sequence of the mouse GPR174 gene (SEQ ID NO: 17).
- FIG. 25 is a diagram showing the base sequence of the rat GPR174 gene (SEQ ID NO: 18).
- FIG. 26 shows the sequences of human, mouse and rat P2Y10 proteins (SEQ ID NOs: 19 to 21).
- FIG. 27 shows the sequences of human, mouse and rat GPR174 proteins (SEQ ID NOs: 22 to 24).
- FIG. 28 is a diagram showing the base sequences of oligonucleotides (SEQ ID NOs: 25 to 30) used as primers in the real-time PCR method of Test Example 4.
- a method for screening a pharmaceutical compound comprising using P2Y10 or GPR174 to evaluate an agonist activity or antagonist activity of a test compound at a lysophosphatidylserine receptor.
- the test compound to be screened is not particularly limited, and examples thereof include organic compounds and inorganic compounds (particularly low molecular compounds), proteins, peptides, and the like.
- the screening is not particularly limited as long as it uses P2Y10 or GPR174.
- a cell expressing a gene encoding P2Y10 or GPR174, a transgenic non-human mammal overexpressing a gene encoding P2Y10 or GPR174, A transgenic non-human mammal expressing a gene encoding human P2Y10 or human GPR174 can be used.
- the cell to be used for screening is not particularly limited, and examples thereof include commonly used known microorganisms such as Escherichia coli or yeast, or known cultured cells such as HUVEC cells. Examples of non-human mammals include mice, rats, rabbits, dogs, cats, monkeys and the like. Screening may also use animal tissues and cells. In that case, administration of the test compound can be performed by allowing the test compound to act on the subject, for example, by including the test compound in a solution or medium in which the tissue or cells are retained.
- the screening method of the present invention can be used for screening a compound having agonistic activity of a test compound against P2Y10 or GPR174. Due to the function of P2Y10 or GPR174, a compound having the agonist activity becomes a candidate compound for a medicament used as a therapeutic or prophylactic agent for diseases such as autoimmune diseases.
- the autoimmune disease may be any of systemic diseases and organ-specific diseases, such as malignant rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, Graves' disease, antiphospholipid antibody syndrome, Examples include Sjogren's syndrome, primary biliary cirrhosis, polymyositis, and autoimmune hepatitis.
- the screening method of the present invention can be performed using a cultured cell that expresses a gene encoding P2Y10 or GPR174, and the use of the cultured cell is preferred in terms of screening efficiency.
- the assay can be performed using calcium response, cAMP production, reporter gene, and the like as indicators.
- the cultured cell is allowed to express a gene encoding a further labeled human epidermal growth factor receptor (EGFR) ligand, and the assay is performed by quantifying the amount of the labeled compound to be cleaved. It can be carried out.
- EGFR human epidermal growth factor receptor
- the labeled EGFR ligand include a fusion protein of an EGFR ligand and alkaline phosphatase (AP).
- AP alkaline phosphatase
- the EGFR ligand to be labeled include transforming growth factor ⁇ (TGF ⁇ ), heparin-binding EGF-like growth factor (HB-EGF), and amphiregulin.
- TGF ⁇ transforming growth factor ⁇
- HB-EGF heparin-binding EGF-like growth factor
- amphiregulin amphiregulin.
- an interleukin-2 production inhibitor or a lymphocyte adhesion inhibitor comprising evaluating the agonist activity of a test compound for a lysophosphatidylserine receptor using P2Y10 or GPR174.
- a method for screening is provided.
- GPR174 is used in the screening method for interleukin 2 production inhibitor
- P2Y10 is used in the screening method for lymphocyte adhesion inhibitor.
- P2Y10 used in the screening method of the present invention is not particularly limited as long as it is a polypeptide having lysophosphatidylserine receptor activity as P2Y10.
- P2Y10 derived from mammals such as humans, mice, and rats is used. . Specific examples include the following polypeptides.
- A a polypeptide comprising an amino acid sequence represented by SEQ ID NOs: 19 to 21;
- B a polypeptide comprising the amino acid sequence represented by SEQ ID NOs: 19 to 21 and having lysophosphatidylserine receptor activity;
- C a polypeptide comprising an amino acid sequence in which 1 to 10 amino acids are deleted, substituted, added and / or inserted in the amino acid sequence represented by SEQ ID NOs: 19 to 21 and having lysophosphatidylserine receptor activity ;
- D an amino acid sequence having a homology with the amino acid sequence represented by SEQ ID NOs: 19 to 21 of 80% or more, preferably 90% or more, more preferably 95% or more, and having lysophosphatidylserine receptor activity Having a polypeptide.
- GPR174 used in the screening method of the present invention is not particularly limited as long as it is a polypeptide having lysophosphatidylserine receptor activity as GPR174.
- GPR174 derived from mammals such as humans, mice and rats is used. . Specific examples include the following polypeptides.
- A a polypeptide comprising an amino acid sequence represented by SEQ ID NOs: 22 to 24;
- B a polypeptide comprising the amino acid sequence represented by SEQ ID NOs: 22 to 24 and having lysophosphatidylserine receptor activity;
- C a polypeptide comprising an amino acid sequence in which 1 to 10 amino acids are deleted, substituted, added and / or inserted in the amino acid sequence represented by SEQ ID NOs: 22 to 24 and having lysophosphatidylserine receptor activity ;
- D an amino acid sequence having a homology with the amino acid sequences represented by SEQ ID NOs: 22 to 24 of 80% or more, preferably 90% or more, more preferably 95% or more, and having lysophosphatidylserine receptor activity Having a polypeptide.
- the numerical value of homology may be a numerical value calculated using a homology search program known to those skilled in the art.
- BLAST Basic Local Alignment Search Tool
- Altschul SF Gish by Altschul et al. W, Miller W, Myers EW, Lipman DJ., J. Mol. Biol., 215: p403-410 (1990)
- Altschyl SF Madden TL, Schaffer AA, Zhang J, Miller W, Lipman DJ, Acids . 25: p3389-3402 (1997)
- amino acid sequence numerical values calculated using default parameters in BLAST2N [Nucleic Acids Res., 25, (3389 (1997); Genome ⁇ ⁇ Res., 7, 649 (1997)] can be mentioned.
- the mutant or homologue when a mutant or homologue of a specific amino acid sequence is used, can be prepared using DNA encoding the amino acid sequence of the mutant or homologue.
- DNA encoding such a mutant or homologue can also be prepared by any method known to those skilled in the art, such as chemical synthesis, genetic engineering techniques, and mutagenesis. Specifically, DNA encoding the sequences described in SEQ ID NOs: 13 to 18 is used, and desired DNA can be obtained by introducing mutations into these DNAs. For example, it can be carried out using a method of bringing DNA into contact with a drug that becomes a mutagen, a method of irradiating ultraviolet rays, a genetic engineering method, or the like.
- Site-directed mutagenesis which is one of the genetic engineering methods, is useful because it can introduce a specific mutation at a specific position.
- Molecular cloning 2nd edition Current Protocols in Molecular. Biology, Nucleic Acids Research, 10, 6487, 1982, Nucleic Acids Research, 12, 9441, 1984, Nucleic Acids Research, 13, 4431, 1985, Nucleic Acids Research, 13, 8749,1985, Proc. Na Sci.Acad. USA, 79, 6409, 1982, Proc. Natl. Acad. Sci. USA, 82, 488, 1985, Gene, 34, 315, 1985, Gene, 102, 67, 1991, etc. Can do.
- lysophosphatidylserine or an analog thereof can be used as a control compound, examples of which include the compounds of the above formulas (I), (Ia) and (Ib), and more specifically And the compounds described in the examples.
- C 1-3 alkyl means a linear, branched, or cyclic alkyl group having 1 to 3 carbon atoms, such as methyl, ethyl, n-propyl, i-propyl, Cyclopropyl is included.
- C 1-6 alkyl means a linear, branched, cyclic or partially cyclic alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl. I-propyl, n-butyl, s-butyl, i-butyl, t-butyl, n-pentyl, 3-methylbutyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, n-hexyl, 4-methylpentyl , 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-ethylbutyl, and 2-ethylbutyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, and the like, for example, C 1-4 alkyl And C 1-3 alkyl and the like are also included.
- C 1-10 alkyl in the definition of the compounds herein, refers to a linear, branched, cyclic or partially cyclic alkyl group having 1 to 10 carbon atoms, e.g., C 1-
- linear, branched, cyclic or partially cyclic represented by C 7 H 15 , C 8 H 17 , C 9 H 19 , and C 10 H 21 These alkyl groups are included.
- C 3-30 alkyl means a linear, branched, cyclic or partially cyclic alkyl group having 7 to 30 carbon atoms, such as C 1-6 alkyl.
- C 1-6 alkylcarbonyl means an alkylcarbonyl group having a C 1-6 alkyl group already defined as the alkyl moiety, such as methylcarbonyl (acetyl), ethylcarbonyl, tert-butylcarbonyl. In addition, C 1-4 alkylcarbonyl and the like are included.
- C 1-6 alkoxy means an alkyloxy group having an alkyl group having 1 to 6 carbon atoms already defined as an alkyl moiety, and includes, for example, methoxy, ethoxy, n-propoxy, i-propoxy N-butoxy, s-butoxy, i-butoxy, t-butoxy, n-pentoxy, 3-methylbutoxy, 2-methylbutoxy, 1-methylbutoxy, 1-ethylpropoxy, n-hexyloxy, 4-methylpen Toxic, 3-methylpentoxy, 2-methylpentoxy, 1-methylpentoxy, 3-ethylbutoxy, cyclopentyloxy, cyclohexyloxy, cyclopropylmethyloxy, etc. are included, for example, C 1-4 alkoxy and C 1 Also included are -3 alkoxy and the like. In this specification, “C 1-4 alkoxy” includes, for example, C 1-3 alkoxy and the like.
- C 1-6 alkoxycarbonyl means an alkoxycarbonyl group having a C 1-6 alkoxy group already defined as an alkoxy moiety, such as methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, C 1-3 alkoxycarbonyl and the like are included.
- C 1-6 alkoxy C 1-6 alkyl in the present specification includes, for example, methoxymethyl, ethoxymethyl, 2-methoxyethyl, 1-methoxyethyl and the like.
- benzyl means a group represented by the formula: C 6 H 5 CH 2 —.
- benzyloxy means a group represented by the formula: C 6 H 5 CH 2 O—.
- benzyloxycarbonyl means a group represented by the formula: C 6 H 5 CH 2 OC ( ⁇ O) —.
- 5-6-membered heteroaryl refers to a monocyclic aromatic heterocyclic group having 5 to 6 ring atoms, including one or more heteroatoms selected from an oxygen atom, a nitrogen atom and a sulfur atom Means.
- Specific examples include pyrrolyl, imidazolyl, pyrazolyl, triazolyl, pyridyl, pyrimidyl, pyridazinyl, furyl, thienyl, oxazolyl, oxadiazolyl, thiazolyl, thiadiazolyl and the like.
- phenylene means a divalent group in which benzene is substituted at two positions, and may be any of 1,2-substituted, 1,3-substituted, and 1,4-substituted. Good.
- 5-6-membered heteroarylene refers to a monocyclic aromatic heterocycle having 5 to 6 ring atoms, including one or more heteroatoms selected from an oxygen atom, a nitrogen atom, and a sulfur atom Means a divalent group substituted at two positions.
- Specific examples of the heterocyclic ring constituting the group include pyrrole, imidazole, pyrazole, triazole, pyridine, pyrimidine, pyridazine, furan, thiophene, oxazole, oxadiazole, thiazole, thiadiazole and the like.
- phenylene or 5-6 membered heteroarylene when one or more phenylene or 5-6 membered heteroarylene is inserted into alkyl, the insertion position of phenylene and 5-6 membered heteroarylene, and the substitution position of phenylene and 5-6 membered heteroarylene are There is no particular limitation.
- the number of inserted phenylene and 5-6 membered heteroarylene is, for example, 1 to 5, preferably 1 to 3.
- —CH 2 — of alkyl when one or more —CH 2 — of alkyl is replaced with —O—, the position of —CH 2 — to be replaced is not particularly limited as long as it can be replaced. Further, phenylene or a 5-6 membered heteroarylene may be inserted between the substituted —O— and —CH 2 — or between —O— and —O—.
- the number of —O— to be replaced is, for example, 1 to 10, preferably 1 to 5, and more preferably 1 to 3.
- the position of the replaced single bond is not particularly limited as long as it can be replaced.
- the double bond may be a cis configuration or a trans configuration.
- the number of double bonds and triple bonds replaced is, for example, 1 to 10, preferably 1 to 5, and more preferably 1 to 3.
- the “pharmaceutically acceptable salt” of the compound of the present invention is not particularly limited as long as it is a salt that can be used as a pharmaceutical product.
- Examples of the salt formed by the compound of the present invention with a base include salts with inorganic bases such as sodium, potassium, magnesium, calcium and aluminum; salts with organic bases such as methylamine, ethylamine and ethanolamine.
- the salt may be an acid addition salt.
- the salt examples include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid and other mineral acids; and formic acid, Examples include acid addition salts with organic acid acids such as acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, and ethanesulfonic acid. .
- the compounds of the present invention also include hydrates, various solvates and crystal polymorphs.
- the atoms (for example, hydrogen atom, carbon atom, oxygen atom, nitrogen atom, sulfur atom and phosphorus atom) contained in the compound of the present invention are isotope atoms other than the most naturally occurring isotopes.
- the isotope atom may be a radioactive conductor atom. That is, according to one aspect of the present invention there is provided a compound of formula (I), (Ia) or (Ib) as defined herein, or a salt thereof, labeled with an isotope atom. .
- labeling with an isotope atom may be, for example, labeling with a radioisotope ( 3 H, 14 C, 32 P, etc.). From the aspect of ease of preparation of the compound, 3 H Labeling is preferred.
- the 3 H-labeled compound of the present invention can be synthesized, for example, by using a 3 H-labeled fatty acid or a derivative thereof.
- the compounds of formula (I), (Ia) and (Ib) are administered as prodrugs and are converted in vivo into active compounds.
- R 5 of formula (I), R 15 of (Ia) and R 25 of (Ib) may be a group forming a phosphate ester. Specific examples thereof include groups described in Journal of Medicinal Chemistry, 2008, 51 (8), 2337, and the like.
- C 1-6 alkyl eg, tert-butyl
- C 1-6 alkoxy C 1- 6 alkyl eg methoxymethyl
- C 1-6 alkylcarbonyloxy C 1-6 alkyl eg pivaloyloxymethyl
- C 1-6 alkoxycarbonyloxy C 1-6 alkyl eg isopropoxycarbonyloxy) Methyl
- optionally substituted phenyl eg, C 1-3 alkoxyphenyl
- optionally substituted benzyl eg, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro, and benzyl which may be substituted by a group of 1 to 3 selected from a halogen atom
- phthalidyl e.g., C 1-6 Good -isobenzofuranone-3-yl optionally substituted by a group having 1 to 4 selected from alkoxy
- di-oxo Lennon yl methyl e
- R 1a and R 2a are each independently selected from C 1-6 alkyl ]
- the compound of the formula (I) can be prepared by condensing the compound 1-1 and the compound 1-2 and then oxidizing.
- the first stage condensation reaction is carried out in a suitable solvent such as methylene chloride, tetrahydrofuran, N, N′-dimethylformamide, toluene, diethyl ether, 1,4-dioxane, etc. (for example, 1H-tetrazole, etc.) )
- This step is not particularly limited, but can be performed, for example, at a reaction temperature of 0 to 70 ° C., preferably 15 to 30 ° C., and a reaction time of, for example, 10 minutes to 2 days, preferably 1 to 2 hours.
- it is desirable that compound 1-1 and compound 1-2 are dissolved in a suitable solvent, mixed and then subjected to azeotropic treatment with a solvent such as toluene or benzene.
- the second stage oxidation step is carried out in a suitable solvent such as, for example, tert-butylhydrochloride in a suitable solvent such as methylene chloride, tetrahydrofuran, N, N′-dimethylformamide, toluene, diethyl ether, 1,4-dioxane.
- a suitable solvent such as methylene chloride, tetrahydrofuran, N, N′-dimethylformamide, toluene, diethyl ether, 1,4-dioxane.
- This step is not particularly limited, but can be performed, for example, at a reaction temperature of 0 to 70 ° C., preferably 15 to 30 ° C., and a reaction time of, for example, 5 minutes to 1 day, preferably 30 minutes to 1 hour.
- the oxidation reaction in the second stage can be performed on the product obtained by post-treating the condensation reaction in the first stage, but can be performed as a one-pot reaction without
- the obtained compound 1-3 has a protecting group, it can be subjected to appropriate deprotection conditions and / or by further substituent substitution to obtain the desired formula (I), (Ia) and (Ib ) Can be obtained.
- the compound of formula (I) can be prepared by acylating compound 2-1 with an acid chloride of compound 2-2.
- the reaction is carried out in a suitable solvent such as methylene chloride, tetrahydrofuran, N, N′-dimethylformamide, toluene, diethyl ether, 1,4-dioxane and the like in the presence of a suitable base (for example, 4-dimethylaminopyridine).
- a suitable base for example, 4-dimethylaminopyridine
- This step is not particularly limited, but can be performed at a reaction temperature of, for example, 0 to 70 ° C., preferably 15 to 30 ° C., and a reaction time of, for example, 10 minutes to 2 days.
- the acylation can also be carried out using a carboxylic acid corresponding to compound 2-2 and a suitable condensing agent (for example, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride).
- a suitable condensing agent for example, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride.
- the obtained compound 2-3 has a protecting group, it can be subjected to appropriate deprotection conditions and / or by further substituent substitution to obtain the desired formula (I), (Ia) and (Ib). ) Can be obtained.
- the pharmaceutical composition of the present invention can be used in various dosage forms such as tablets, capsules, powders, granules, pills, solutions, emulsions, suspensions, solutions, spirits, syrups for oral administration.
- parenteral preparations include, for example, injections such as subcutaneous injections, intravenous injections, intramuscular injections, intraperitoneal injections; transdermal administration or patches , Ointments or lotions; sublingual and buccal patches for buccal administration; and aerosols for nasal administration, but not limited thereto. These preparations can be produced by known methods usually used in the preparation process.
- the compounds of formula (I), (Ia) and (Ib) are preferably administered as parenteral agents.
- the pharmaceutical composition may contain various commonly used components, such as one or more pharmaceutically acceptable excipients, disintegrants, diluents, lubricants, flavoring agents, and coloring agents. , Sweeteners, flavoring agents, suspending agents, wetting agents, emulsifying agents, dispersing agents, adjuvants, preservatives, buffering agents, binders, stabilizers, coating agents and the like.
- the pharmaceutical composition of the present invention may be in a sustained or sustained release dosage form.
- the dosage of the therapeutic agent, prophylactic agent, or pharmaceutical composition of the present invention can be appropriately selected depending on the administration route, the patient's body shape, age, physical condition, degree of disease, elapsed time after onset, etc.
- the pharmaceutical composition can comprise a therapeutically effective amount and / or a prophylactically effective amount of a compound of formula (I), (Ia), or (Ib) above.
- the compound of the above formula (I), formula (Ia) or formula (Ib) can be generally used at a dose of 1 to 10000 mg / day / adult.
- the pharmaceutical composition may be administered in a single dose or multiple doses, such as immunosuppressants (cyclosporine, tacrolimus, sirolimus, methotrexate, azathioprine, etc.), steroidal anti-inflammatory drugs (hydrocortisone, prednisolone, dexamethasone) Etc.), non-steroidal anti-inflammatory drugs (such as loxoprofen sodium, indomethacin, diclofenac sodium), or antibody drugs (such as infliximab, adalimumab, tocilizumab, sertolizumab pegol, etanercept) You can also.
- immunosuppressants cyclosporine, tacrolimus, sirolimus, methotrexate, azathioprine, etc.
- steroidal anti-inflammatory drugs hydrocortisone, prednisolone, dexamethasone
- non-steroidal anti-inflammatory drugs such as loxoprofen sodium
- the therapeutic agent or prophylactic agent of the present invention may contain a conventionally known coloring agent, preservative, flavoring, flavoring agent, coating agent, antioxidant, vitamin, amino acid, peptide, protein, and mineral content (iron, zinc, if necessary). , Magnesium, iodine, etc.).
- the therapeutic agent or prophylactic agent of the present invention is in a form suitable for pharmaceutical compositions, functional foods, health foods, beverages, supplements, etc., such as granules (including dry syrup), capsules (soft capsules, hard capsules). , Prepared in the form of various solid preparations such as tablets (including chewables), powders (powder), pills, or liquid preparations for internal use (including liquids, suspensions, syrups) May be.
- the therapeutic agent or prophylactic agent of the present invention can be used as it is as a pharmaceutical composition, functional food, health food, supplement or the like.
- additives for formulation for example, excipients, lubricants, binders, disintegrants, fluidizers, dispersants, wetting agents, preservatives, thickeners, pH adjusters, colorants, Examples include flavoring agents, surfactants, and solubilizing agents.
- thickeners such as pectin, xanthan gum, and guar gum, can be mix
- thickeners such as pectin, xanthan gum, and guar gum
- it can also be set as a coating tablet using a coating agent, or it can also be set as a paste-form glue. Furthermore, even if it is a case where it prepares in another form, what is necessary is just to follow the conventional method.
- Reagents and data measurement reagents are available from Sigma-Aldrich Chemical Co. Those purchased from Tokyo Chemical Industry, Wako Pure Chemicals, and Kanto Chemical were used without further purification. 1 H- and 13 C-NMR were measured using a BRUKER AVANCE 400 spectrometer (400 MHz), and the chemical shift was deuterated chloroform (7.26 ppm ( 1 H-NMR), 77.00 ppm ( 13 C-NMR)). In ppm. 31 P-NMR chemical shifts are expressed in ppm relative to phosphoric acid in water (85% w / w, 0.00 ppm).
- Mass spectrometry was measured in the BRUKER microTOF-05 spectrometer (ESI-TOF) or a SHIMADZU AXIMA-TOF (MALDI-TOF) positive and negative ion mode.
- Silica gel used for column chromatography was purchased from Kanto Chemical. Elemental analysis was performed using a Yanaco MT-6 CHN CORDER spectrometer.
- 1,3-Diisopropylcarbodiimide (6.325 g, 50.1 mmol) and CuCl (50.8 mg, 0.513 mmol) were dissolved in anhydrous tBuOH (4.556 g, 59.9 mmol), and at room temperature under argon atmosphere for 3 days Stir. A small portion of the reaction mixture was taken and the completion of the reaction was confirmed by 1 H-NMR. Anhydrous CH 2 Cl 2 (25 ml) and polyvinylpyrrolidone (1.004 g) were added to the reaction mixture, stirred for 15 minutes and filtered through Celite®. The filtrate was concentrated to give the title compound (8.18 g, 40.47 mmol, 81%, green oil). The obtained compound was used as an esterification reagent without further purification.
- Step 2 Preparation of N-tert-butoxycarbonylserine tert-butyl ester
- L-serine (1.025 g, 9.756 mmol) was dissolved in a mixture of 1N NaOH aqueous solution (1N, 10 ml), H 2 O (10 ml) and dioxane (20 ml), and the mixture was stirred at 0 ° C.
- Di-tert-butyl dicarbonate (3.192 g, 14.63 mmol) was added to the solution at 0 ° C., and the mixture was stirred as it was for 10 minutes and then at room temperature for 24 hours.
- 5% KHSO 4 aqueous solution was added to the reaction mixture to bring the pH to 3 and extracted with AcOEt (150 ml ⁇ 3). The organic layers were combined, washed with brine and dried over Na 2 SO 4 .
- N-Boc serine was distilled off to give N-Boc serine as a crude product (2.228 g, colorless oil).
- Step 3 Preparation of (2S) -3-[(diisopropylamino) -tert-butoxyphosphinooxy] -2- (tert-butoxycarbonylamino) propionic acid tert-butyl ester
- 1,3-propanediol (623.3 mg, 8.192 mmol) and pyridine (0.2 ml) were dissolved in CH 2 Cl 2 (15 ml) and oleoyl chloride (623.4 mg in CH 2 Cl 2 (5 ml). , 2.072 mmol) was added dropwise at 0 ° C.
- the reaction mixture was stirred for 24 hours and quenched with 5% KHSO 4 aqueous solution (15 ml).
- the organic layer was separated, washed with 5% aqueous KHSO 4 solution and brine, dried over Na 2 SO 4 and concentrated.
- Step 5 Preparation of (2S) -2- (tert-butoxycarbonylamino) -3-[(3-oleoyloxypropoxy) -tert-butoxy-phosphoryloxy] propionic acid tertbutyl ester
- Example 9 In the production of Example 9, the following compounds were used as synthetic intermediates.
- Example 13 In the production of Example 13, the following compounds were used as synthetic intermediates.
- Step 1 Preparation of 8-azidooctanoic acid N 3 — (CH 2 ) 7 COOH
- DMF DMF
- NaN 3 5.76 g, 88.60 mmol
- KHSO 4 aqueous solution 1N, 50 ml
- hexane 100 ml ⁇ 3
- Step 2 Preparation of 8- (4-heptyl-1,2,3-triazol-1-yl) octanoic acid
- 1,3-propanediol 324.4 mg, 4.263 mmol
- 6- ⁇ 4-hexyl- (2Z) -2-buten-1-yloxy ⁇ hexanoic acid 300.2 mg, 1.048 mmol
- 4-dimethyl Aminopyridine (14.3 mg, 0.119 mmol) was dissolved in CH 2 Cl 2 (10 ml) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (212.5 mg, 1.109 mmol) was added to 0. The mixture was added at ° C and stirred at room temperature for 19 hours.
- Example 16 In the production of Example 16, the following compounds were used as synthetic intermediates.
- Step 1 3-tert-butyldimethylsilyloxy-1-propanol TBSO- (CH 2 ) 3 —OH NaH (790.9 mg, 19.77 mmol) was suspended in THF (3 ml) and stirred at 0 ° C. To the suspension was added dropwise a solution of 1,3-propanediol (1000.4 mg, 13.15 mmol) in THF (3 ml) and stirred at 0 ° C. for 30 minutes. A solution of tert-butyldimethylsilyl chloride (2.3841 g, 5.82 mmol) in THF (4 ml) was added dropwise and stirred at room temperature for 22.5 hours.
- Step 3 3-tert-butyldimethylsilyloxy-propionic acid TBSO- (CH 2 ) 2 —COOH 3-tert-butyldimethylsilyloxy-1-propanal (1.0438 g, 5.542 mmol) and 2-methyl-2-butene (1.7191 g, 24.51 mmol) were dissolved in t BuOH (15 ml) and H A solution of NaH 2 PO 4 .2H 2 O (867.8 mg, 5.562 mmol) and NaClO 2 (2.0425 g, 22.58 mmol) in 2 O (5 ml) was added and stirred at room temperature for 1 hour.
- Step 1 Preparation of N-tert-butoxycarbonyl-L-allo-threonine tert-butyl ester
- Step 2 (2S, 3S) -3-[(Diisopropylamino) -tert-butoxyphosphinooxy] -2- (tert-butoxycarbonylamino) -3-methyl-propionic acid tert-butyl ester
- Step 1 Preparation of (R) -4- (4-methoxyphenoxy) methyl-2,2-dimethyl-1,3-dioxolane
- Step 5 (2S, 3S) -2-tert-butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) propoxy) -tert-butoxy [Phosphoryloxy] butyric acid tert-butyl ester
- tert-butyl hydroperoxide (TBHP) decane solution (5.0-6.0 M, 0.251 ml, 1.255 mmol) was added at room temperature, and the mixture was stirred at room temperature for 1.5 hours. did.
- Step 7 (2S, 3S) -2-tert-Butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3-hydroxypropoxy) -tert-butoxyphosphoryloxy] butyric acid tert -Preparation of butyl esters
- Step 8 Preparation of (2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid
- reaction mixture was stirred at room temperature for 3.5 hours, tert-butyl hydroperoxide (TBHP) decane solution (5.0-6.0 M, 0.431 ml, 2.155 mmol) was added at room temperature, and the mixture was further stirred for 1.5 hours. did.
- TBHP tert-butyl hydroperoxide
- Step 7 Preparation of (2S) -2-amino-3-[((2R) -2-hydroxy-3- ⁇ 3- (2-heptyloxyphenyl) propionyloxy ⁇ propoxy) -hydroxyphosphoryloxy] propionic acid
- Step 1 Cloning of P2Y10 and GPR174 genes
- Full-length cDNAs of human, mouse, and rat P2Y10 and GPR174 genes were obtained by PCR using HUVEC cells, mouse tail genome, and rat tail genome as templates.
- the oligonucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 1-6 is used as a forward primer
- the oligonucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 7-12 is used as a reverse primer Using.
- a base sequence containing a restriction enzyme KpnKI recognition site is added to the 5 ′ end of each of the forward primers.
- a base sequence including a restriction enzyme Sma I recognition site is added to the 3 ′ ′ end of human P2Y10 of the reverse primer, and restriction enzyme EcoR is added to the 3 ′ ′ end of human GPR174, mouse and rat P2Y10 and GPR174.
- Each nucleotide sequence including a V recognition site is added.
- PrimeSTARPrimHS # R010Q, 010TAKARA
- the base sequences represented by SEQ ID NOs: 13 to 15 have open reading frames (O R F) of 984 bases, 987 bases, and 987 bases in human, mouse, and rat, respectively, and are predicted from this O R F.
- the amino acid sequences (328 amino acids, 329 amino acids, and 329 amino acids) were as shown in SEQ ID NOs: 19 to 21, respectively.
- the base sequences represented by SEQ ID NOs: 16 to 18 have ORFs of 1002 bases, 1008 bases and 1008 bases in human, mouse and rat, respectively, and amino acid sequences predicted from these ORFs (each 334 Amino acids, 336 amino acids, 336 amino acids) were as shown in the amino acid sequences represented by SEQ ID NOs: 22 to 24.
- Step 2 Evaluation of agonist activity by TGF ⁇ cleavage assay HEK293 cells are suspended in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal calf serum (FCS) at 2.0x10 5 cells / mL, and 4 mL in a 60 mm dish. Seeded one by one. After culturing for 24 hours in the presence of 5% CO 2 , various expression vectors were transfected using LipofectAMINE 2000 (Invitrogen). 1 ⁇ g of AP-tagged TGF ⁇ plasmid vector and 1 ⁇ g of mouse P2Y10 or GPR174 plasmid vector were used per 60 mm dish.
- DMEM Dulbecco's modified Eagle's medium
- FCS fetal calf serum
- AP-labeled TGF ⁇ is a protein in which human placental alkaline phosphatase is fused to the N-terminal side of membrane-bound pro-TGF ⁇ , and the AP-labeled TGF ⁇ plasmid vector is Tokumaru et al., J Cell Biol 151, 209-220 ( 2000). Twenty-four hours after transfection, the cells were detached using trypsin / EDTA, resuspended in a culture solution to 2.0 ⁇ 10 4 cells / well, and seeded in a 96-well plate. When stimulating with a compound, Hanks balanced salt solution (HBSS, containing 5 mM HEPES) was used for resuspension.
- HBSS Hanks balanced salt solution
- test compounds were added, and the mixture was further allowed to stand for 1 hour in the presence of 5% CO 2 .
- the 96-well plate was centrifuged (190 ⁇ g, 3 minutes), and 80 ⁇ L of the supernatant was transferred to another 96-well plate.
- 80 ⁇ L of a reaction buffer 40 mM Tris-HCl (pH 9.5)
- p-NPP p-nitrophenyl phosphate
- OD405 was measured with a microplate reader (background), heated at 37 ° C. for 1 hour, and then measured again.
- the AP activity was calculated by applying a value obtained by subtracting the background from the second absorbance to the following equation.
- Example 34 LPS (18: 1)
- Example 1 deoxy-LPS (18: 1)
- Example 19 LPalloT (18: 1)
- deoxy-LPS (18: 1) is an agonist having selectivity for P2Y10
- LPalloT (18: 1) is a selectivity for GPR174.
- Emax maximum response value
- MRL-lpr / lpr mice (hereinafter also referred to as MRL / lpr mice, 8-9 weeks old, rabbit) were purchased from Japan SLC Co., Ltd.
- the compound of Example 34 (LPS (18: 1), purchased from Avanti Polar Lipids, 858143C) was taken into a silicon-coated glass tube (Asahi Techno Glass, 9831-1207) with a microsyringe and dried to nitrogen, 0.01% (w / v) BSA-PBS was added, vortexed, and sonicated with a bath sonicator for 2 minutes to prepare 10 mM LPS.
- a small osmotic pump (Alzet, model 2006) was soaked in physiological saline (Otsuka raw food injection) and incubated at 37 ° C for 60 hours. Thereafter, each osmotic pump was filled with 200 ⁇ L of LPS solution and 0.01% (w / v) BSA-PBS using a 1 mL syringe (TERUMO, SS-01T) and the attached 27G needle. After administration of Nembutal anesthetic to Balb / c mice at 100 mg / kg, a small osmotic pump was implanted vertically into the abdominal cavity of the mouse, and the peritoneum and abdominal wall were sutured.
- MRL / lpr mice ( ⁇ , 9 weeks old) were filled with the compound of Example 34 (LPS (18: 1), 10 mM) (small discharge rate 0.15 ⁇ L / hr, 42 days) ) Was implanted into the abdominal cavity to provide a continuous supply of LPS (18: 1).
- LPS low-density polypeptide
- VetScan HM2 fully automated blood cell device
- Measurement of the amount of autoantibodies was performed by the following procedure.
- a 96-well assay plate was coated with a poly-L-lysine solution (50 ⁇ L / well) and allowed to stand at room temperature for 8 hours to coat the well bottom.
- 100 ⁇ l / well of 1 ng / ⁇ L DNA solution was added and incubated at 37 ° C. for 8 hours to bind to the plate.
- 180 ⁇ L / well of blocking buffer was added, and blocking was performed at room temperature for 2 hours.
- a serum sample diluted with assay buffer was added at 50 ⁇ L / well and allowed to stand at room temperature for 2 hours. Then wash 3 times with TTBS using Skan Washer 400 (Molecular Devices), add 50 ⁇ L / well of biotinylated anti-mouse IgG (H + L) (Vector, BA2001) diluted 2000 times with Assay buffer, and react for 2 hours. I let you.
- kidney paraffin sections were prepared and immunocomplexes were stained with anti-mouse IgG antibodies. Paraffin sections were deparaffinized by sequentially immersing them in Xylen (3 times), 100% (3 times), 90%, 70%, and 50% EtOH for 3 minutes each, and then washed with running water for 5 minutes. Thereafter, the slide was immersed in 3% H 2 O 2 for 30 minutes to inactivate the tissue's endogenous peroxidase, and washed again with running water for 5 minutes. Using a pressure cooker, the slide glass was immersed in 1 mM EDTA to activate the antigen, and then endogenous biotin was blocked using an Avidin / Biotin Blocking kit (VECTOR, SP-2001).
- VECTOR Avidin / Biotin Blocking kit
- the plate was washed with PBS, DAPI was added and nuclear staining was performed for 5 minutes, and then the excess solution was washed off with PBS and encapsulated using an encapsulating agent, Aqueous Mounting Medium, PermaFluor (Thermo, TA-030-FM).
- an encapsulating agent Aqueous Mounting Medium, PermaFluor (Thermo, TA-030-FM).
- the small osmotic pump is used in the same procedure as in Test Example 2. Implanted in the abdominal cavity of mice. After 4 days, the animals were fasted for 12 hours, and concanavalin A (ConA) was intravenously injected at 20 mg / kg. After a certain time, blood was collected to prepare serum. After perfusion with PBS, the liver was removed and paraffin blocks and sections were prepared. Alanine transaminase (ALT) in serum was measured using transaminase CII test Wako. Paraffin sections were stained with hematoxylin and eosin and observed using an optical microscope.
- ConA concanavalin A
- spleen cells activated with ConA were collected, and total RNA was extracted using GenElute Mammalian Total RNA kit (SIGMA-ALDRICH). Further, cDNA was obtained by reverse transcription using a high capacity cDNA RT kit (Applied Biosystems) according to a conventional method. cDNA, a primer for measuring a target gene, and SYBR Green PCR Master Mix (Applied Biosystems) were mixed, and PCR products were quantified by real-time PCR using 7300 Real-Time PCR System. Oligonucleotides shown in SEQ ID NOs: 25 to 28 were used as primers. Further, GAPDH primers used as control genes are shown in SEQ ID NOs: 29 and 30. The expression levels of P2Y10 receptor and GPR174 receptor are shown in FIGS. It was confirmed that the expression level of any receptor increased upon stimulation with concanavalin A.
- Spleen cells activated with ConA were collected, washed twice with PBS, suspended in PBS containing 1 ⁇ M CMFDA, and fluorescently labeled at 37 ° C. for 1 hour. Further, the spleen cells were washed twice with PBS and suspended in RPMI1640 medium (containing 0.01% bovine serum albumin (BSA)). A 96-well black plate was treated with 2.5 ⁇ g / ml recombinant mouse ICAM Fc at 4 ° C. for 24 hours, and blocked with PBS containing 0.5% BSA for 30 minutes. 96 well palte was washed 3 times with PBS, and spleen cells were seeded at 2 ⁇ 10 5 cells / well.
- BSA bovine serum albumin
- LPS (18: 1), deoxy LPS (18: 1), and LPalloT (18: 1) were used.
- the test results are shown in FIG. From the test results, it was confirmed that LPS (18: 1) and deoxy LPS (18: 1) inhibit adhesion of spleen cells to ICAM-1, but LPalloT (18: 1) does not inhibit this adhesion ( Compound final concentration: 10 ⁇ M).
- mice Male, body weight 20-25 g
- RPMI1640 medium 100 mU / ml penicillin, 100 ⁇ g
- streptomycin 2 mM L-glutamine, 10% FBS
- Stimulation was performed in the presence of 1 ⁇ g / ml ConA, and the cells were cultured at 37 ° C. for 60 hours.
- Spleen cells activated with ConA were collected, suspended in RPMI1640 medium (containing 0.01% BSA), and seeded on a 96-well plate at 5 ⁇ 10 5 cells / well.
- Activated spleen cells in the presence of test compounds were stimulated at 37 ° C. with 1 ⁇ / ml ConA to promote IL-2 production.
- the culture supernatant was collected from the 96 well plate.
- the concentration of IL-2 in the supernatant was determined by sandwich ELISA using Mouse IL-2 Matched Antibody Pairs for ELISA (eBioscience, BMS601MST). The measured value was expressed as mean value ⁇ standard error after several repeated experiments using spleen cells collected from different mouse individuals.
- LPS (18: 1), deoxy-LPS (18: 1), and LPalloT (18: 1) were used.
- the test results are shown in FIG. From the test results, LPS (18: 1) and LPalloT (18: 1) inhibit IL-2 production in spleen cells in a concentration-dependent manner, but deoxy-LPS (18: 1) inhibits IL-2 production. It was confirmed not to.
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Abstract
The present invention provides a screening method for a preventive or therapeutic agent for an autoimmune disease, comprising evaluating agonist activity or antagonist activity of a test compound on a lysophosphatidylserine receptor using P2Y10 or GPR174.
Description
本発明は、自己免疫治療薬として有用な化合物のスクリーニング方法に関する。また本発明は、自己免疫疾患治療薬に関する。さらに本発明は、リゾホスファチジルセリンおよび新規なその誘導体に関する。
The present invention relates to a screening method for compounds useful as autoimmune therapeutic agents. The present invention also relates to a therapeutic agent for autoimmune diseases. The invention further relates to lysophosphatidylserine and novel derivatives thereof.
リゾリン脂質はアシル基を1本有するリン脂質の総称である。リゾリン脂質は、細胞膜を構成するジアシルリン脂質に比べ疎水性が低下しており、容易に細胞膜から遊離することができるという性質を有している。いくつかのリゾリン脂質は、細胞間あるいは膜間のシグナル分子として機能し、生体内で重要な役割を持っていることが明らかとなっており、従来から、組織障害などの炎症反応が起こる際、生体膜のリン脂質が加水分解されリゾリン脂質が産生されることが知られている(特許文献1:特開2007-267601号)。リゾリン脂質の一種であるリゾホスファチジルセリン(LysoPS)はマスト細胞の脱顆粒に起因する急性炎症に関与することが知られている(非特許文献1および2)。LysoPS受容体として、Gタンパク質共役受容体の中から、LPS1(GPR34)、LPS2(P2Y10)、LPS3(A630033H20Rik)、およびLPS4(GPR174)が同定されている(非特許文献3)。このうちLPS1はマスト細胞の脱顆粒反応を誘導または増強するシグナル伝達に関与し、アレルギー性疾患および慢性炎症疾患の処置のための標的となりうるとの報告がされている(特許文献1)。また特定のリゾホスファチジルセリン誘導体(リゾホスファチジルスレオニン)がマスト細胞の脱顆粒反応を強力に促進することが知られている(特許文献2)。
Lysophospholipid is a general term for phospholipids having one acyl group. The lysophospholipid has a property that it is less hydrophobic than the diacylphospholipid constituting the cell membrane and can be easily released from the cell membrane. Some lysophospholipids function as signal molecules between cells or between membranes and have been found to have an important role in vivo. Traditionally, when inflammatory reactions such as tissue damage occur, It is known that phospholipids in biological membranes are hydrolyzed to produce lysophospholipids (Patent Document 1: Japanese Patent Application Laid-Open No. 2007-267601). It is known that lysophosphatidylserine (LysoPS), which is a kind of lysophospholipid, is involved in acute inflammation caused by mast cell degranulation (Non-patent Documents 1 and 2). Among the G protein coupled receptors, LPS 1 (GPR34), LPS 2 (P2Y10), LPS 3 (A630033H20Rik), and LPS 4 (GPR174) have been identified as LysoPS receptors (Non-patent Document 3). Among them, it has been reported that LPS 1 is involved in signal transduction that induces or enhances degranulation of mast cells and can be a target for the treatment of allergic diseases and chronic inflammatory diseases (Patent Document 1). In addition, it is known that a specific lysophosphatidylserine derivative (lysophosphatidylthreonine) strongly promotes the degranulation reaction of mast cells (Patent Document 2).
自己免疫疾患は、本来は異物に対する防御機構である免疫系が過剰に反応し、自分自身の正常な細胞や組織に対してまで攻撃を加えてしまうことで症状を来す疾患の総称であり、全身にわたり影響が及ぶ全身性自己免疫疾患と、特定の臓器だけが影響を受ける臓器特異的疾患の2種類に大別される。自己免疫疾患は、一般的に慢性疾患、難治性疾患となることが多く、そのいくつかは厚生労働省特定疾患研究対象疾患に指定されている。自己免疫疾患の処置方法については多くの研究がなされており、例えば、炎症に関与するサイトカイン特異的抗体を用いた自己免疫疾患に起因する慢性炎症の処置方法(特許文献2)、および病原性自己抗体の中和による疾患の治療方法(特許文献3)などが報告されている。しかし、自己免疫疾患の原因は解明されていない点が多く、有効な治療法は多くの自己免疫疾患でまだ確立されていない。自己免疫疾患に対しては免疫系を抑制する薬剤や炎症を和らげる抗炎症薬(ステロイドまたは非ステロイド薬)などが第一選択剤として用いられている。
Autoimmune disease is a general term for diseases that cause symptoms when the immune system, which is originally a defense mechanism against foreign substances, reacts excessively and attacks even normal cells and tissues of itself. There are two broad categories: systemic autoimmune diseases that affect the whole body and organ-specific diseases that affect only specific organs. In general, autoimmune diseases often become chronic diseases or intractable diseases, and some of them are designated as diseases for which specific diseases are researched by the Ministry of Health, Labor and Welfare. Much research has been conducted on methods for treating autoimmune diseases, for example, methods for treating chronic inflammation caused by autoimmune diseases using cytokine-specific antibodies involved in inflammation (Patent Document 2), and pathogenic self. A method for treating diseases by neutralizing antibodies (Patent Document 3) has been reported. However, many causes of autoimmune diseases have not been elucidated, and effective treatments have not yet been established for many autoimmune diseases. For autoimmune diseases, drugs that suppress the immune system and anti-inflammatory drugs (steroids or non-steroid drugs) that relieve inflammation are used as the first selection agent.
本発明は、新たな自己免疫疾患の処置方法を提供することを目的とする。さらに本発明は、自己免疫疾患の処置に有用な化合物のアッセイ方法を提供することである。
An object of the present invention is to provide a new treatment method for autoimmune diseases. Furthermore, the present invention is to provide a method for assaying compounds useful for the treatment of autoimmune diseases.
本発明者らは、上記課題を達成するために鋭意研究を進めたところ、LPS2(P2Y10)およびLPS4(GPR174)のアゴニスト活性を有する化合物に自己免疫疾患に対する治療および予防効果を見いだし、本発明を完成させた。
The inventors of the present invention have made extensive studies in order to achieve the above-mentioned problems. As a result, the compounds having agonist activity of LPS 2 (P2Y10) and LPS 4 (GPR174) have been found to have therapeutic and preventive effects on autoimmune diseases. Completed the invention.
本発明の1つの側面によれば、以下のスクリーニング方法が提供される。
(1)P2Y10またはGPR174を使用して、リゾホスファチジルセリン受容体への被検化合物のアゴニスト活性またはアンタゴニスト活性を評価することを含む、自己免疫疾患の治療剤または予防剤のスクリーニング方法。
(2)アゴニスト活性を評価する、上記(1)に記載の方法。
(3)インターロイキン2産生抑制剤をスクリーニングするための、上記(1)または(2)に記載の方法。
(4)リンパ球接着抑制剤をスクリーニングするための、上記(1)または(2)に記載の方法。
(5)P2Y10またはGPR174をコードする遺伝子を発現する細胞を使用する、上記(1)~(4)のいずれかに記載の方法。
(6)細胞がさらに標識化されたヒト上皮増殖因子受容体(EGFR)リガンドをコードする遺伝子を発現する、上記(5)に記載の方法。
(7)標識化されたEGFRリガンドが、EGFRリガンドとアルカリフォスファターゼ(AP)の融合タンパク質である、上記(6)に記載の方法。
(8)標識化されたEGFRリガンドが、標識化されたトランスフォーミング増殖因子α(TGFα)である、上記(6)または(7)に記載の方法。
(9)対照化合物としてリゾホスファチジルセリンまたはその類縁体を使用する、上記(1)~(8)のいずれかに記載の方法。 According to one aspect of the present invention, the following screening method is provided.
(1) A screening method for a therapeutic or prophylactic agent for an autoimmune disease, comprising using P2Y10 or GPR174 to evaluate the agonist activity or antagonist activity of a test compound for a lysophosphatidylserine receptor.
(2) The method according to (1) above, wherein the agonist activity is evaluated.
(3) The method according to (1) or (2) above for screening for aninterleukin 2 production inhibitor.
(4) The method according to (1) or (2) above for screening for a lymphocyte adhesion inhibitor.
(5) The method according to any one of (1) to (4) above, wherein a cell expressing a gene encoding P2Y10 or GPR174 is used.
(6) The method according to (5) above, wherein the cell further expresses a gene encoding a labeled human epidermal growth factor receptor (EGFR) ligand.
(7) The method according to (6) above, wherein the labeled EGFR ligand is a fusion protein of an EGFR ligand and alkaline phosphatase (AP).
(8) The method according to (6) or (7) above, wherein the labeled EGFR ligand is labeled transforming growth factor α (TGFα).
(9) The method according to any one of (1) to (8) above, wherein lysophosphatidylserine or an analog thereof is used as a control compound.
(1)P2Y10またはGPR174を使用して、リゾホスファチジルセリン受容体への被検化合物のアゴニスト活性またはアンタゴニスト活性を評価することを含む、自己免疫疾患の治療剤または予防剤のスクリーニング方法。
(2)アゴニスト活性を評価する、上記(1)に記載の方法。
(3)インターロイキン2産生抑制剤をスクリーニングするための、上記(1)または(2)に記載の方法。
(4)リンパ球接着抑制剤をスクリーニングするための、上記(1)または(2)に記載の方法。
(5)P2Y10またはGPR174をコードする遺伝子を発現する細胞を使用する、上記(1)~(4)のいずれかに記載の方法。
(6)細胞がさらに標識化されたヒト上皮増殖因子受容体(EGFR)リガンドをコードする遺伝子を発現する、上記(5)に記載の方法。
(7)標識化されたEGFRリガンドが、EGFRリガンドとアルカリフォスファターゼ(AP)の融合タンパク質である、上記(6)に記載の方法。
(8)標識化されたEGFRリガンドが、標識化されたトランスフォーミング増殖因子α(TGFα)である、上記(6)または(7)に記載の方法。
(9)対照化合物としてリゾホスファチジルセリンまたはその類縁体を使用する、上記(1)~(8)のいずれかに記載の方法。 According to one aspect of the present invention, the following screening method is provided.
(1) A screening method for a therapeutic or prophylactic agent for an autoimmune disease, comprising using P2Y10 or GPR174 to evaluate the agonist activity or antagonist activity of a test compound for a lysophosphatidylserine receptor.
(2) The method according to (1) above, wherein the agonist activity is evaluated.
(3) The method according to (1) or (2) above for screening for an
(4) The method according to (1) or (2) above for screening for a lymphocyte adhesion inhibitor.
(5) The method according to any one of (1) to (4) above, wherein a cell expressing a gene encoding P2Y10 or GPR174 is used.
(6) The method according to (5) above, wherein the cell further expresses a gene encoding a labeled human epidermal growth factor receptor (EGFR) ligand.
(7) The method according to (6) above, wherein the labeled EGFR ligand is a fusion protein of an EGFR ligand and alkaline phosphatase (AP).
(8) The method according to (6) or (7) above, wherein the labeled EGFR ligand is labeled transforming growth factor α (TGFα).
(9) The method according to any one of (1) to (8) above, wherein lysophosphatidylserine or an analog thereof is used as a control compound.
(10)自己免疫疾患が、悪性関節リウマチ、全身エリテマトーデス、多発性硬化症、重症筋無力症、バセドウ病、抗リン脂質抗体症候群、シェーグレン症候群、原発性胆汁性肝硬変、多発性筋炎、および自己免疫性肝炎から選択される疾患である、上記(1)~(9)のいずれかに記載の方法。 本発明者らは、LPS2(P2Y10)およびLPS4(GPR174)のアゴニスト活性を有する化合物を見いだした。すなわち本発明の別の側面によれば、以下の化合物が提供される。
(10) Autoimmune diseases are malignant rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, Graves' disease, antiphospholipid antibody syndrome, Sjogren's syndrome, primary biliary cirrhosis, polymyositis, and autoimmunity The method according to any one of (1) to (9) above, which is a disease selected from sexual hepatitis. The inventors have found compounds having agonist activity of LPS 2 (P2Y10) and LPS 4 (GPR174). That is, according to another aspect of the present invention, the following compounds are provided.
(11)式(Ia):
(11) Formula (Ia):
[式中、A1は下式:
[Wherein A 1 is the following formula:
で表される基であり;
R11は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、またはベンジルであり;
R12およびR13は、それぞれ独立に、水素原子、C1-6アルキル、ホルミル、C1-6アルキルカルボニル、C1-6アルコキシカルボニル、C1-6アルコキシC1-6アルキル、またはベンジルオキシカルボニルから選択され;
R15は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、C1-6アルキルカルボニルオキシC1-6アルキル、C1-6アルコキシカルボニルオキシC1-6アルキル、フェニル、ベンジル、フタリジル、ジオキソレノンイルメチル、またはフリルメチルであり;
pは、2~6から選択される整数であり;
Y1は、直接結合、または-C(=O)-であり;
R16は、C3-30アルキルであり、ここでアルキルには1以上のフェニレンまたは5-6員ヘテロアリーレンが挿入されていてもよく、アルキルの1以上の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1以上の単結合は、二重結合または三重結合に置き換えられていてもよく、アルキルの末端はフェニルまたは5-6員ヘテロアリールにより置換されていてもよく、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシおよびハロゲン原子から選択される1以上の置換基により置換されていてもよく;
上記のベンジル、ベンジルオキシ、フェニル、フタリジル、ジオキソレノンイルメチル、フリルメチル、フェニレン、およびヘテロアリーレンは、C1-6アルキル、C1-6アルコキシ、ヒドロキシ、C1-6アルキルカルボニルオキシ、ニトロ、フェニルおよびハロゲン原子から選択される1以上の置換基により置換されていてもよい]
で表される化合物、または医薬として許容なその塩。 A group represented by:
R 11 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, or benzyl;
R 12 and R 13 are each independently a hydrogen atom, C 1-6 alkyl, formyl, C 1-6 alkylcarbonyl, C 1-6 alkoxycarbonyl, C 1-6 alkoxy C 1-6 alkyl, or benzyloxy Selected from carbonyl;
R 15 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, C 1-6 alkylcarbonyloxy C 1-6 alkyl, C 1-6 alkoxycarbonyloxy C 1-6 alkyl, Phenyl, benzyl, phthalidyl, dioxolenoneylmethyl, or furylmethyl;
p is an integer selected from 2 to 6;
Y 1 is a direct bond or —C (═O) —;
R 16 is C 3-30 alkyl, wherein one or more phenylene or 5-6 membered heteroarylene may be inserted in the alkyl, and one or more —CH 2 — of alkyl is —O— and One or more single bonds between carbon atoms may be replaced by double bonds or triple bonds, and the alkyl ends may be substituted by phenyl or 5-6 membered heteroaryl. Well, the phenyl or heteroaryl may be substituted with one or more substituents selected from C 1-10 alkyl, C 1-10 alkoxy and a halogen atom;
The above benzyl, benzyloxy, phenyl, phthalidyl, dioxolenoneylmethyl, furylmethyl, phenylene, and heteroarylene are C 1-6 alkyl, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro May be substituted with one or more substituents selected from phenyl and halogen atoms]
Or a pharmaceutically acceptable salt thereof.
R11は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、またはベンジルであり;
R12およびR13は、それぞれ独立に、水素原子、C1-6アルキル、ホルミル、C1-6アルキルカルボニル、C1-6アルコキシカルボニル、C1-6アルコキシC1-6アルキル、またはベンジルオキシカルボニルから選択され;
R15は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、C1-6アルキルカルボニルオキシC1-6アルキル、C1-6アルコキシカルボニルオキシC1-6アルキル、フェニル、ベンジル、フタリジル、ジオキソレノンイルメチル、またはフリルメチルであり;
pは、2~6から選択される整数であり;
Y1は、直接結合、または-C(=O)-であり;
R16は、C3-30アルキルであり、ここでアルキルには1以上のフェニレンまたは5-6員ヘテロアリーレンが挿入されていてもよく、アルキルの1以上の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1以上の単結合は、二重結合または三重結合に置き換えられていてもよく、アルキルの末端はフェニルまたは5-6員ヘテロアリールにより置換されていてもよく、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシおよびハロゲン原子から選択される1以上の置換基により置換されていてもよく;
上記のベンジル、ベンジルオキシ、フェニル、フタリジル、ジオキソレノンイルメチル、フリルメチル、フェニレン、およびヘテロアリーレンは、C1-6アルキル、C1-6アルコキシ、ヒドロキシ、C1-6アルキルカルボニルオキシ、ニトロ、フェニルおよびハロゲン原子から選択される1以上の置換基により置換されていてもよい]
で表される化合物、または医薬として許容なその塩。 A group represented by:
R 11 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, or benzyl;
R 12 and R 13 are each independently a hydrogen atom, C 1-6 alkyl, formyl, C 1-6 alkylcarbonyl, C 1-6 alkoxycarbonyl, C 1-6 alkoxy C 1-6 alkyl, or benzyloxy Selected from carbonyl;
R 15 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, C 1-6 alkylcarbonyloxy C 1-6 alkyl, C 1-6 alkoxycarbonyloxy C 1-6 alkyl, Phenyl, benzyl, phthalidyl, dioxolenoneylmethyl, or furylmethyl;
p is an integer selected from 2 to 6;
Y 1 is a direct bond or —C (═O) —;
R 16 is C 3-30 alkyl, wherein one or more phenylene or 5-6 membered heteroarylene may be inserted in the alkyl, and one or more —CH 2 — of alkyl is —O— and One or more single bonds between carbon atoms may be replaced by double bonds or triple bonds, and the alkyl ends may be substituted by phenyl or 5-6 membered heteroaryl. Well, the phenyl or heteroaryl may be substituted with one or more substituents selected from C 1-10 alkyl, C 1-10 alkoxy and a halogen atom;
The above benzyl, benzyloxy, phenyl, phthalidyl, dioxolenoneylmethyl, furylmethyl, phenylene, and heteroarylene are C 1-6 alkyl, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro May be substituted with one or more substituents selected from phenyl and halogen atoms]
Or a pharmaceutically acceptable salt thereof.
(12)R16が、C10-24アルキル(ここでアルキルの1以上の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1以上の単結合は、二重結合または三重結合に置き換えられていてもよい)、または式:-(CH2)r-Q1から選択され;
rは、2~20から選択される整数であり;
Q1は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-10アルキル、フェニルC1-10アルコキシ、5-6員ヘテロアリールC1-10アルキル、または5-6員ヘテロアリールC1-10アルコキシにより置換されていてもよい、上記(11)に記載の化合物、または医薬として許容なその塩。 (12) R 16 is C 10-24 alkyl (wherein one or more —CH 2 — of alkyl may be replaced by —O—, and one or more single bonds between carbon atoms are double bonds) Or may be replaced by a triple bond), or selected from the formula: — (CH 2 ) r —Q 1 ;
r is an integer selected from 2 to 20;
Q 1 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-10 alkyl, phenyl C 1-10 alkoxy, 5- The compound according to the above (11), or a pharmaceutically acceptable salt thereof, which may be substituted with 6-membered heteroaryl C 1-10 alkyl or 5-6-membered heteroaryl C 1-10 alkoxy.
rは、2~20から選択される整数であり;
Q1は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-10アルキル、フェニルC1-10アルコキシ、5-6員ヘテロアリールC1-10アルキル、または5-6員ヘテロアリールC1-10アルコキシにより置換されていてもよい、上記(11)に記載の化合物、または医薬として許容なその塩。 (12) R 16 is C 10-24 alkyl (wherein one or more —CH 2 — of alkyl may be replaced by —O—, and one or more single bonds between carbon atoms are double bonds) Or may be replaced by a triple bond), or selected from the formula: — (CH 2 ) r —Q 1 ;
r is an integer selected from 2 to 20;
Q 1 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-10 alkyl, phenyl C 1-10 alkoxy, 5- The compound according to the above (11), or a pharmaceutically acceptable salt thereof, which may be substituted with 6-membered heteroaryl C 1-10 alkyl or 5-6-membered heteroaryl C 1-10 alkoxy.
(13)R16が、C12-20アルキル(ここでアルキルの1~3の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1つの単結合は、二重結合に置き換えられていてもよい)、または式:-(CH2)r-Q1から選択され;
rは、2~7から選択される整数であり;
Q1は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-3アルキル、フェニルC1-3アルコキシ、5-6員ヘテロアリールC1-3アルキル、または5-6員ヘテロアリールC1-3アルコキシにより置換されていてもよい、上記(12)に記載の化合物、または医薬として許容なその塩。 (13) R 16 is C 12-20 alkyl (whereinalkyl 1 to 3 —CH 2 — may be replaced by —O—, and one single bond between carbon atoms is a double bond) Or is selected from the formula: — (CH 2 ) r —Q 1 ;
r is an integer selected from 2 to 7;
Q 1 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5- The compound according to the above (12), or a pharmaceutically acceptable salt thereof, which may be substituted by 6-membered heteroaryl C 1-3 alkyl or 5-6-membered heteroaryl C 1-3 alkoxy.
rは、2~7から選択される整数であり;
Q1は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-3アルキル、フェニルC1-3アルコキシ、5-6員ヘテロアリールC1-3アルキル、または5-6員ヘテロアリールC1-3アルコキシにより置換されていてもよい、上記(12)に記載の化合物、または医薬として許容なその塩。 (13) R 16 is C 12-20 alkyl (wherein
r is an integer selected from 2 to 7;
Q 1 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5- The compound according to the above (12), or a pharmaceutically acceptable salt thereof, which may be substituted by 6-membered heteroaryl C 1-3 alkyl or 5-6-membered heteroaryl C 1-3 alkoxy.
さらに本発明の別の側面によれば、以下の化合物が提供される。
Furthermore, according to another aspect of the present invention, the following compounds are provided.
(14)式(Ib):
(14) Formula (Ib):
R21は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、またはベンジルであり;
R22およびR23は、それぞれ独立に、水素原子、C1-6アルキル、ホルミル、C1-6アルキルカルボニル、C1-6アルコキシカルボニル、C1-6アルコキシC1-6アルキル、またはベンジルオキシカルボニルから選択され;
R24は、水素原子またはC1-3アルキルであり;
R25は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、C1-6アルキルカルボニルオキシC1-6アルキル、C1-6アルコキシカルボニルオキシC1-6アルキル、フェニル、ベンジル、フタリジル、ジオキソレノンイルメチル、またはフリルメチルであり;
X2は、水素原子、ハロゲン原子、ヒドロキシ、C1-6アルコキシ、ホルミルオキシ、C1-6アルキルカルボニルオキシ、またはベンジルオキシであり;
sは、0~4から選択される整数であり;
Y2は、-O-、-C(=O)O-、-OC(=O)-、-C(=O)NR27-、または-NR27C(=O)-であり;
R26は、式:-(CH2)t-Q2であり;
tは、2~20から選択される整数であり;
Q2は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-10アルキル、フェニルC1-10アルコキシ、5-6員ヘテロアリールC1-10アルキル、または5-6員ヘテロアリールC1-10アルコキシにより置換されていてもよく;
R27は、水素原子、またはC1-6アルキルであり;
さらに上記のベンジル、ベンジルオキシ、フェニル、フタリジル、ジオキソレノンイルメチル、およびフリルメチルは、C1-6アルキル、C1-6アルコキシ、ヒドロキシ、C1-6アルキルカルボニルオキシ、ニトロ、フェニルおよびハロゲン原子から選択される1以上の置換基により置換されていてもよい]
で表される化合物、または医薬として許容なその塩。
R 21 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, or benzyl;
R 22 and R 23 are each independently a hydrogen atom, C 1-6 alkyl, formyl, C 1-6 alkylcarbonyl, C 1-6 alkoxycarbonyl, C 1-6 alkoxy C 1-6 alkyl, or benzyloxy Selected from carbonyl;
R 24 is a hydrogen atom or C 1-3 alkyl;
R 25 represents a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, C 1-6 alkylcarbonyloxy C 1-6 alkyl, C 1-6 alkoxycarbonyloxy C 1-6 alkyl, Phenyl, benzyl, phthalidyl, dioxolenoneylmethyl, or furylmethyl;
X 2 is a hydrogen atom, a halogen atom, hydroxy, C 1-6 alkoxy, formyloxy, C 1-6 alkylcarbonyloxy, or benzyloxy;
s is an integer selected from 0 to 4;
Y 2 is —O—, —C (═O) O—, —OC (═O) —, —C (═O) NR 27 —, or —NR 27 C (═O) —;
R 26 is of the formula: — (CH 2 ) t —Q 2 ;
t is an integer selected from 2 to 20;
Q 2 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-10 alkyl, phenyl C 1-10 alkoxy, 5- Optionally substituted by 6-membered heteroaryl C 1-10 alkyl, or 5-6-membered heteroaryl C 1-10 alkoxy;
R 27 is a hydrogen atom or C 1-6 alkyl;
Further, the above benzyl, benzyloxy, phenyl, phthalidyl, dioxolenoneylmethyl and furylmethyl are C 1-6 alkyl, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro, phenyl and halogen. Optionally substituted by one or more substituents selected from atoms]
Or a pharmaceutically acceptable salt thereof.
(15)A2が下式:
(15) A 2 is the following formula:
(16)R26が、C12-20アルキル(ここでアルキルの1~3の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1つの単結合は、二重結合に置き換えられていてもよい)、または式:-(CH2)s-Q2から選択され;
sは、2~7から選択される整数であり;
Q2は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-3アルキル、フェニルC1-3アルコキシ、5-6員ヘテロアリールC1-3アルキル、または5-6員ヘテロアリールC1-3アルコキシにより置換されていてもよい、上記(14)または(15)に記載の化合物、または医薬として許容なその塩。 (16) R 26 is C 12-20 alkyl (wherein 1 to 3 —CH 2 — of alkyl may be replaced by —O—, and one single bond between carbon atoms is a double bond) Or is selected from the formula: — (CH 2 ) s —Q 2 ;
s is an integer selected from 2 to 7;
Q 2 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5- The compound according to the above (14) or (15), or a pharmaceutically acceptable salt thereof, which may be substituted with 6-membered heteroaryl C 1-3 alkyl, or 5-6-membered heteroaryl C 1-3 alkoxy .
sは、2~7から選択される整数であり;
Q2は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-3アルキル、フェニルC1-3アルコキシ、5-6員ヘテロアリールC1-3アルキル、または5-6員ヘテロアリールC1-3アルコキシにより置換されていてもよい、上記(14)または(15)に記載の化合物、または医薬として許容なその塩。 (16) R 26 is C 12-20 alkyl (wherein 1 to 3 —CH 2 — of alkyl may be replaced by —O—, and one single bond between carbon atoms is a double bond) Or is selected from the formula: — (CH 2 ) s —Q 2 ;
s is an integer selected from 2 to 7;
Q 2 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5- The compound according to the above (14) or (15), or a pharmaceutically acceptable salt thereof, which may be substituted with 6-membered heteroaryl C 1-3 alkyl, or 5-6-membered heteroaryl C 1-3 alkoxy .
また本発明者らは、リゾホスファチジルセリン誘導体に自己免疫疾患に対する治療および予防効果を見いだし、本発明を完成させた。すなわち本発明のさらに別の側面によれば、以下の治療剤または予防剤が提供される。
In addition, the present inventors have found that lysophosphatidylserine derivatives have therapeutic and preventive effects on autoimmune diseases, and have completed the present invention. That is, according to still another aspect of the present invention, the following therapeutic or preventive agent is provided.
(17)式(I):
(17) Formula (I):
R1は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、またはベンジルであり;
R2およびR3は、それぞれ独立に、水素原子、C1-6アルキル、ホルミル、C1-6アルキルカルボニル、C1-6アルコキシカルボニル、C1-6アルコキシC1-6アルキル、またはベンジルオキシカルボニルから選択され;
R4は、水素原子またはC1-3アルキルであり;
R5は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、C1-6アルキルカルボニルオキシC1-6アルキル、C1-6アルコキシカルボニルオキシC1-6アルキル、フェニル、ベンジル、フタリジル、ジオキソレノンイルメチル、またはフリルメチルであり;
Xは、水素原子、ハロゲン原子、ヒドロキシ、C1-6アルコキシ、ホルミルオキシ、C1-6アルキルカルボニルオキシ、またはベンジルオキシであり;
nは、0~4から選択される整数であり;
Yは、-O-、-C(=O)O-、-OC(=O)-、-C(=O)NR7-、または-NR7C(=O)-であり;
R6は、C3-30アルキルであり、ここでアルキルには1以上のフェニレンまたは5-6員ヘテロアリーレンが挿入されていてもよく、アルキルの1以上の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1以上の単結合は、二重結合または三重結合に置き換えられていてもよく、アルキルの末端はフェニルまたは5-6員ヘテロアリールにより置換されていてもよく、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシおよびハロゲン原子から選択される1以上の置換基により置換されていてもよく;
R7は、水素原子、またはC1-6アルキルであり;
上記のベンジル、ベンジルオキシ、フェニル、フタリジル、ジオキソレノンイルメチル、フリルメチル、フェニレン、およびヘテロアリーレンは、C1-6アルキル、C1-6アルコキシ、ヒドロキシ、C1-6アルキルカルボニルオキシ、ニトロ、フェニルおよびハロゲン原子から選択される1以上の置換基により置換されていてもよい]
で表される化合物、または医薬として許容なその塩を含有する、自己免疫疾患の治療剤または予防剤。
R 1 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, or benzyl;
R 2 and R 3 are each independently a hydrogen atom, C 1-6 alkyl, formyl, C 1-6 alkylcarbonyl, C 1-6 alkoxycarbonyl, C 1-6 alkoxy C 1-6 alkyl, or benzyloxy Selected from carbonyl;
R 4 is a hydrogen atom or C 1-3 alkyl;
R 5 represents a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, C 1-6 alkylcarbonyloxy C 1-6 alkyl, C 1-6 alkoxycarbonyloxy C 1-6 alkyl, Phenyl, benzyl, phthalidyl, dioxolenoneylmethyl, or furylmethyl;
X is a hydrogen atom, a halogen atom, hydroxy, C 1-6 alkoxy, formyloxy, C 1-6 alkylcarbonyloxy, or benzyloxy;
n is an integer selected from 0 to 4;
Y is —O—, —C (═O) O—, —OC (═O) —, —C (═O) NR 7 —, or —NR 7 C (═O) —;
R 6 is C 3-30 alkyl, wherein one or more phenylene or 5-6 membered heteroarylene may be inserted in the alkyl, and one or more —CH 2 — of alkyl is —O— and One or more single bonds between carbon atoms may be replaced by double bonds or triple bonds, and the alkyl ends may be substituted by phenyl or 5-6 membered heteroaryl. Well, the phenyl or heteroaryl may be substituted with one or more substituents selected from C 1-10 alkyl, C 1-10 alkoxy and a halogen atom;
R 7 is a hydrogen atom or C 1-6 alkyl;
The above benzyl, benzyloxy, phenyl, phthalidyl, dioxolenoneylmethyl, furylmethyl, phenylene, and heteroarylene are C 1-6 alkyl, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro May be substituted with one or more substituents selected from phenyl and halogen atoms]
The therapeutic agent or preventive agent of an autoimmune disease containing the compound represented by these, or its pharmaceutically acceptable salt.
(18)Aが下式:
(18) A is the following formula:
(19)R6が、C10-24アルキル(ここでアルキルの1以上の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1以上の単結合は、二重結合または三重結合に置き換えられていてもよい)、または式:-(CH2)m-Qから選択され;
mは、2~20から選択される整数であり;
Qは、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-10アルキル、フェニルC1-10アルコキシ、5-6員ヘテロアリールC1-10アルキル、または5-6員ヘテロアリールC1-10アルコキシにより置換されていてもよい、上記(17)または(18)に記載の治療剤または予防剤。 (19) R 6 is C 10-24 alkyl (wherein one or more —CH 2 — of alkyl may be replaced by —O—, and one or more single bonds between carbon atoms are double bonds) Or may be replaced by a triple bond), or selected from the formula: — (CH 2 ) m —Q;
m is an integer selected from 2 to 20;
Q is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-10 alkyl, phenyl C 1-10 alkoxy, 5-6 The therapeutic or prophylactic agent according to the above (17) or (18), which may be substituted with a member heteroaryl C 1-10 alkyl or a 5-6 membered heteroaryl C 1-10 alkoxy.
mは、2~20から選択される整数であり;
Qは、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-10アルキル、フェニルC1-10アルコキシ、5-6員ヘテロアリールC1-10アルキル、または5-6員ヘテロアリールC1-10アルコキシにより置換されていてもよい、上記(17)または(18)に記載の治療剤または予防剤。 (19) R 6 is C 10-24 alkyl (wherein one or more —CH 2 — of alkyl may be replaced by —O—, and one or more single bonds between carbon atoms are double bonds) Or may be replaced by a triple bond), or selected from the formula: — (CH 2 ) m —Q;
m is an integer selected from 2 to 20;
Q is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-10 alkyl, phenyl C 1-10 alkoxy, 5-6 The therapeutic or prophylactic agent according to the above (17) or (18), which may be substituted with a member heteroaryl C 1-10 alkyl or a 5-6 membered heteroaryl C 1-10 alkoxy.
(20)R6が、C12-20アルキル(ここでアルキルの1~3の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1つの単結合は、二重結合に置き換えられていてもよい)、または式:-(CH2)m-Qから選択され;
mは、2~7から選択される整数であり;
Qは、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-3アルキル、フェニルC1-3アルコキシ、5-6員ヘテロアリールC1-3アルキル、または5-6員ヘテロアリールC1-3アルコキシにより置換されていてもよい、上記(19)に記載の治療剤または予防剤。 (20) R 6 is C 12-20 alkyl (wherein the alkyl 1-3 —CH 2 — may be replaced by —O—, and one single bond between carbon atoms is a double bond) Or may be selected from the formula: — (CH 2 ) m —Q;
m is an integer selected from 2 to 7;
Q is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5-6 The therapeutic or prophylactic agent according to the above (19), which may be substituted with a member heteroaryl C 1-3 alkyl or a 5-6 membered heteroaryl C 1-3 alkoxy.
mは、2~7から選択される整数であり;
Qは、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-3アルキル、フェニルC1-3アルコキシ、5-6員ヘテロアリールC1-3アルキル、または5-6員ヘテロアリールC1-3アルコキシにより置換されていてもよい、上記(19)に記載の治療剤または予防剤。 (20) R 6 is C 12-20 alkyl (wherein the alkyl 1-3 —CH 2 — may be replaced by —O—, and one single bond between carbon atoms is a double bond) Or may be selected from the formula: — (CH 2 ) m —Q;
m is an integer selected from 2 to 7;
Q is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5-6 The therapeutic or prophylactic agent according to the above (19), which may be substituted with a member heteroaryl C 1-3 alkyl or a 5-6 membered heteroaryl C 1-3 alkoxy.
(21)(2S)-2-アミノ-3-[(3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-バセノイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-エライドイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-パルミトレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-ミリストイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-ラウロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-ヘキサデシルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(2-オレイルオキシエトキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-オレイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(4-オレイルオキシブトキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(5-オレイルオキシペントキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[3-{8-(4-ヘプチル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[3-{8-(4-ヘキシル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[6-{4-ヘキシルオキシ-(2Z)-2-ブテン-1-イルオキシ}ヘキサノイルオキシ]プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[2-(オレイルオキシカルボニル)エトキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[3-(オレイルオキシカルボニル)プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[4-(オレイルオキシカルボニル)ブトキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S,3S)-2-アミノ-3-[(3-オレオイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[{3-((9E)-オクタデセノイルオキシ)プロピル}-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[(3-ステアロイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[(3-パルミトレオイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[(3-パルミトイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸;
(2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(2-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(3-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(4-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(4-ベンジルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;および
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
から選択される化合物、または医薬として許容なその塩を含有する、上記(17)~(20)のいずれかに記載の治療剤または予防剤。 (21) (2S) -2-Amino-3-[(3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3-[(3-basenoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-Elideyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-palmitoleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-myristoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-lauroyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-hexadecyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(2-oleyloxyethoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-oleyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(4-oleyloxybutoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(5-oleyloxypentoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [3- {8- (4-heptyl-1,2,3-triazol-1-yl) octanoyloxy} propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [3- {8- (4-hexyl-1,2,3-triazol-1-yl) octanoyloxy} propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [6- {4-hexyloxy- (2Z) -2-buten-1-yloxy} hexanoyloxy] propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [2- (oleyloxycarbonyl) ethoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [3- (oleyloxycarbonyl) propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [4- (oleyloxycarbonyl) butoxy-hydroxyphosphoryloxy] propionic acid;
(2S, 3S) -2-Amino-3-[(3-oleoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[{3-((9E) -octadecenoyloxy) propyl} -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-Amino-3-[(3-stearoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[(3-palmitoleoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-Amino-3-[(3-palmitoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid;
(2S) -2-amino-3-[((2S) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2S) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2S) -2-hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (2-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (3-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (4-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (4-benzyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid; and (2S) -2-amino-3-[((2R) -2-Hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid or a pharmaceutically acceptable salt thereof, the treatment according to any one of (17) to (20) above Or preventive agent.
(2S)-2-アミノ-3-[(3-バセノイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-エライドイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-パルミトレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-ミリストイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-ラウロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-ヘキサデシルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(2-オレイルオキシエトキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-オレイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(4-オレイルオキシブトキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(5-オレイルオキシペントキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[3-{8-(4-ヘプチル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[3-{8-(4-ヘキシル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[6-{4-ヘキシルオキシ-(2Z)-2-ブテン-1-イルオキシ}ヘキサノイルオキシ]プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[2-(オレイルオキシカルボニル)エトキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[3-(オレイルオキシカルボニル)プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[4-(オレイルオキシカルボニル)ブトキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S,3S)-2-アミノ-3-[(3-オレオイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[{3-((9E)-オクタデセノイルオキシ)プロピル}-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[(3-ステアロイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[(3-パルミトレオイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[(3-パルミトイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸;
(2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(2-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(3-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(4-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(4-ベンジルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;および
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
から選択される化合物、または医薬として許容なその塩を含有する、上記(17)~(20)のいずれかに記載の治療剤または予防剤。 (21) (2S) -2-Amino-3-[(3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3-[(3-basenoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-Elideyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-palmitoleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-myristoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-lauroyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-hexadecyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(2-oleyloxyethoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-oleyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(4-oleyloxybutoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(5-oleyloxypentoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [3- {8- (4-heptyl-1,2,3-triazol-1-yl) octanoyloxy} propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [3- {8- (4-hexyl-1,2,3-triazol-1-yl) octanoyloxy} propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [6- {4-hexyloxy- (2Z) -2-buten-1-yloxy} hexanoyloxy] propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [2- (oleyloxycarbonyl) ethoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [3- (oleyloxycarbonyl) propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [4- (oleyloxycarbonyl) butoxy-hydroxyphosphoryloxy] propionic acid;
(2S, 3S) -2-Amino-3-[(3-oleoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[{3-((9E) -octadecenoyloxy) propyl} -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-Amino-3-[(3-stearoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[(3-palmitoleoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-Amino-3-[(3-palmitoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid;
(2S) -2-amino-3-[((2S) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2S) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2S) -2-hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (2-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (3-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (4-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (4-benzyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid; and (2S) -2-amino-3-[((2R) -2-Hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid or a pharmaceutically acceptable salt thereof, the treatment according to any one of (17) to (20) above Or preventive agent.
(22)自己免疫疾患が、悪性関節リウマチ、全身エリテマトーデス、多発性硬化症、重症筋無力症、バセドウ病、抗リン脂質抗体症候群、シェーグレン症候群、原発性胆汁性肝硬変、多発性筋炎、および自己免疫性肝炎から選択される疾患である、上記(17)~(21)のいずれかに記載の治療剤または予防剤。
(22) The autoimmune diseases are malignant rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, Graves' disease, antiphospholipid antibody syndrome, Sjogren's syndrome, primary biliary cirrhosis, polymyositis, and autoimmunity The therapeutic or prophylactic agent according to any one of (17) to (21) above, which is a disease selected from sexual hepatitis.
さらに本発明の別の側面によれば、上記(11)~(16)のいずれかに記載の化合物、もしくは上記(17)~(21)のいずれかにおいて定義される式(I)の化合物、または医薬として許容なその塩を含有する、自己免疫疾患を治療または予防するための医薬組成物が提供される。
Furthermore, according to another aspect of the present invention, the compound according to any one of (11) to (16) above, or the compound of the formula (I) defined in any one of (17) to (21) above, Or the pharmaceutical composition for treating or preventing an autoimmune disease containing the pharmaceutically acceptable salt is provided.
さらに本発明の別の側面によれば、上記(11)~(16)のいずれかに記載の化合物、もしくは上記(17)~(21)のいずれかにおいて定義される式(I)の化合物、または医薬として許容なその塩を含有する、インターロイキン2産生抑制剤が提供される。
Furthermore, according to another aspect of the present invention, the compound according to any one of (11) to (16) above, or the compound of the formula (I) defined in any one of (17) to (21) above, Alternatively, an interleukin 2 production inhibitor containing a pharmaceutically acceptable salt thereof is provided.
さらに本発明の別の側面によれば、上記(11)~(16)のいずれかに記載の化合物、もしくは上記(17)~(21)のいずれかにおいて定義される式(I)の化合物、または医薬として許容なその塩を含有する、リンパ球接着抑制剤が提供される。
Furthermore, according to another aspect of the present invention, the compound according to any one of (11) to (16) above, or the compound of the formula (I) defined in any one of (17) to (21) above, Alternatively, a lymphocyte adhesion inhibitor containing a pharmaceutically acceptable salt thereof is provided.
さらに本発明の別の側面によれば、上記(11)~(16)のいずれかに記載の化合物、もしくは上記(17)~(21)のいずれかにおいて定義される式(I)の化合物、または医薬として許容なその塩を患者に投与することを含む、自己免疫疾患の治療方法または予防方法が提供される。
Furthermore, according to another aspect of the present invention, the compound according to any one of (11) to (16) above, or the compound of the formula (I) defined in any one of (17) to (21) above, Alternatively, a method of treating or preventing autoimmune disease comprising administering to a patient a pharmaceutically acceptable salt thereof.
本発明によれば、新たな自己免疫疾患の処置方法となる自己免疫疾患の治療剤または予防剤が提供される。さらに本発明は、自己免疫疾患の処置に有用な化合物の効率なスクリーニング方法が提供される。
According to the present invention, there is provided a therapeutic or prophylactic agent for autoimmune diseases, which is a new method for treating autoimmune diseases. Furthermore, the present invention provides an efficient screening method for compounds useful for the treatment of autoimmune diseases.
以下、本発明を更に具体的に説明する。
Hereinafter, the present invention will be described more specifically.
本発明の1つの側面によれば、P2Y10またはGPR174を使用して、リゾホスファチジルセリン受容体への被検化合物のアゴニスト活性またはアンタゴニスト活性を評価することを含む、医薬化合物のスクリーニング方法が提供される。スクリーニング対象となる被検化合物としては特に限定されず、有機化合物および無機化合物(特に、低分子化合物)、タンパク質、ペプチドなどが例として挙げられる。
According to one aspect of the present invention, there is provided a method for screening a pharmaceutical compound, comprising using P2Y10 or GPR174 to evaluate an agonist activity or antagonist activity of a test compound at a lysophosphatidylserine receptor. . The test compound to be screened is not particularly limited, and examples thereof include organic compounds and inorganic compounds (particularly low molecular compounds), proteins, peptides, and the like.
スクリーニングはP2Y10またはGPR174を使用するものであれば特に限定されず、例えば、P2Y10またはGPR174をコードする遺伝子を発現する細胞、P2Y10またはGPR174をコードする遺伝子を過剰に発現するトランスジェニック非ヒト哺乳動物、ヒトP2Y10またはヒトGPR174をコードする遺伝子を発現するトランスジェニック非ヒト哺乳動物などを用いて行うことができる。ここで、スクリーニングに使用する細胞としては、特には限定されないが、通常使用されている公知の微生物、例えば大腸菌、または酵母、あるいは公知の培養細胞、例えば、HUVEC細胞などが挙げられる。また、非ヒト哺乳動物の例としては、マウス、ラット、ウサギ、イヌ、ネコ、サルなどが挙げられる。また、スクリーニングは動物の組織および細胞を使用してもよい。その場合、被検化合物の投与は、例えば、組織または細胞が保持される溶液または培地などに被検化合物を含ませることなどにより、対象に被検化合物を作用させることにより行うことができる。
The screening is not particularly limited as long as it uses P2Y10 or GPR174. For example, a cell expressing a gene encoding P2Y10 or GPR174, a transgenic non-human mammal overexpressing a gene encoding P2Y10 or GPR174, A transgenic non-human mammal expressing a gene encoding human P2Y10 or human GPR174 can be used. Here, the cell to be used for screening is not particularly limited, and examples thereof include commonly used known microorganisms such as Escherichia coli or yeast, or known cultured cells such as HUVEC cells. Examples of non-human mammals include mice, rats, rabbits, dogs, cats, monkeys and the like. Screening may also use animal tissues and cells. In that case, administration of the test compound can be performed by allowing the test compound to act on the subject, for example, by including the test compound in a solution or medium in which the tissue or cells are retained.
本発明のスクリーニング方法は、P2Y10またはGPR174への被検化合物のアゴニスト活性を有する化合物のスクリーニングに利用することができる。P2Y10またはGPR174の機能から、当該アゴニスト活性を有する化合物は、自己免疫疾患などの疾患の治療剤または予防剤として使用される医薬の候補化合物となる。自己免疫疾患としては、全身性疾患および臓器特異性疾患の何れであってもよく、例えば、悪性関節リウマチ、全身エリテマトーデス、多発性硬化症、重症筋無力症、バセドウ病、抗リン脂質抗体症候群、シェーグレン症候群、原発性胆汁性肝硬変、多発性筋炎、および自己免疫性肝炎などが挙げられる。
The screening method of the present invention can be used for screening a compound having agonistic activity of a test compound against P2Y10 or GPR174. Due to the function of P2Y10 or GPR174, a compound having the agonist activity becomes a candidate compound for a medicament used as a therapeutic or prophylactic agent for diseases such as autoimmune diseases. The autoimmune disease may be any of systemic diseases and organ-specific diseases, such as malignant rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, Graves' disease, antiphospholipid antibody syndrome, Examples include Sjogren's syndrome, primary biliary cirrhosis, polymyositis, and autoimmune hepatitis.
本発明の1つの態様において、本発明のスクリーニング方法はP2Y10またはGPR174をコードする遺伝子を発現する培養細胞を使用して行うことができ、培養細胞の使用は、スクリーニングの効率の点で好ましい。また、P2Y10またはGPR174はGタンパク質共役型受容体であることから、カルシウム応答、cAMP産生、レポーター遺伝子などを指標としてアッセイを行うことができる。本発明の1つの態様として、前記培養細胞にさらに標識化されたヒト上皮増殖因子受容体(EGFR)リガンドをコードする遺伝子を発現させて、切断される標識化合物の量を定量することによりアッセイを行うことができる。ここで、標識されたEGFRリガンドの具体例としては、EGFRリガンドとアルカリフォスファターゼ(AP)の融合タンパク質などが挙げられる。また、標識化されるEGFRリガンドとしては、トランスフォーミング増殖因子α(TGFα)、ヘパリン結合性EGF様増殖因子(HB-EGF)、およびアンフィレグリン (Amphiregulin)などが挙げられる。標識されたEGFRリガンドを用いるアッセイ方法は、例えば、Tokumaru et al., J Cell Biol 151, 209-220 (2000);石黒ら、脂質生化学研究 52, 221-224, (2010)を参照して行うことができる。
In one embodiment of the present invention, the screening method of the present invention can be performed using a cultured cell that expresses a gene encoding P2Y10 or GPR174, and the use of the cultured cell is preferred in terms of screening efficiency. In addition, since P2Y10 or GPR174 is a G protein-coupled receptor, the assay can be performed using calcium response, cAMP production, reporter gene, and the like as indicators. In one embodiment of the present invention, the cultured cell is allowed to express a gene encoding a further labeled human epidermal growth factor receptor (EGFR) ligand, and the assay is performed by quantifying the amount of the labeled compound to be cleaved. It can be carried out. Here, specific examples of the labeled EGFR ligand include a fusion protein of an EGFR ligand and alkaline phosphatase (AP). Examples of the EGFR ligand to be labeled include transforming growth factor α (TGFα), heparin-binding EGF-like growth factor (HB-EGF), and amphiregulin. For assay methods using labeled EGFR ligands, see, for example, Tokumaru et al., J Cell Biol 151, 209-220 (2000); Ishiguro et al., Lipid Biochemical Research 52, 221-224, (2010). It can be carried out.
本発明の1つの側面によれば、P2Y10またはGPR174を使用して、リゾホスファチジルセリン受容体への被検化合物のアゴニスト活性を評価することを含む、インターロイキン2産生抑制剤またはリンパ球接着抑制剤をスクリーニングするための方法が提供される。好ましくは、インターロイキン2産生抑制剤のスクリーニング方法において、GPR174が使用され、リンパ球接着抑制剤のスクリーニング方法においてP2Y10が使用される。
According to one aspect of the present invention, an interleukin-2 production inhibitor or a lymphocyte adhesion inhibitor comprising evaluating the agonist activity of a test compound for a lysophosphatidylserine receptor using P2Y10 or GPR174. A method for screening is provided. Preferably, GPR174 is used in the screening method for interleukin 2 production inhibitor, and P2Y10 is used in the screening method for lymphocyte adhesion inhibitor.
本発明のスクリーニング方法において使用されるP2Y10は、P2Y10としてのリゾホスファチジルセリン受容体活性を有するポリペプチドであれば特に限定されす、例えば、ヒト、マウス、ラットなどの哺乳類由来のP2Y10が使用される。具体的な例としては、以下のポリペプチドが挙げられる。
P2Y10 used in the screening method of the present invention is not particularly limited as long as it is a polypeptide having lysophosphatidylserine receptor activity as P2Y10. For example, P2Y10 derived from mammals such as humans, mice, and rats is used. . Specific examples include the following polypeptides.
(a)配列番号19~21で表されるアミノ酸配列からなるポリペプチド;
(b)配列番号19~21で表されるアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド;
(c)配列番号19~21で表されるアミノ酸配列において1~10のアミノ酸が欠失、置換、付加、および/または挿入されたアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド;
(d)配列番号19~21で表されるアミノ酸配列との相同性が80%以上、好ましくは90%以上、より好ましくは95%以上であるアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド。 (A) a polypeptide comprising an amino acid sequence represented by SEQ ID NOs: 19 to 21;
(B) a polypeptide comprising the amino acid sequence represented by SEQ ID NOs: 19 to 21 and having lysophosphatidylserine receptor activity;
(C) a polypeptide comprising an amino acid sequence in which 1 to 10 amino acids are deleted, substituted, added and / or inserted in the amino acid sequence represented by SEQ ID NOs: 19 to 21 and having lysophosphatidylserine receptor activity ;
(D) an amino acid sequence having a homology with the amino acid sequence represented by SEQ ID NOs: 19 to 21 of 80% or more, preferably 90% or more, more preferably 95% or more, and having lysophosphatidylserine receptor activity Having a polypeptide.
(b)配列番号19~21で表されるアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド;
(c)配列番号19~21で表されるアミノ酸配列において1~10のアミノ酸が欠失、置換、付加、および/または挿入されたアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド;
(d)配列番号19~21で表されるアミノ酸配列との相同性が80%以上、好ましくは90%以上、より好ましくは95%以上であるアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド。 (A) a polypeptide comprising an amino acid sequence represented by SEQ ID NOs: 19 to 21;
(B) a polypeptide comprising the amino acid sequence represented by SEQ ID NOs: 19 to 21 and having lysophosphatidylserine receptor activity;
(C) a polypeptide comprising an amino acid sequence in which 1 to 10 amino acids are deleted, substituted, added and / or inserted in the amino acid sequence represented by SEQ ID NOs: 19 to 21 and having lysophosphatidylserine receptor activity ;
(D) an amino acid sequence having a homology with the amino acid sequence represented by SEQ ID NOs: 19 to 21 of 80% or more, preferably 90% or more, more preferably 95% or more, and having lysophosphatidylserine receptor activity Having a polypeptide.
本発明のスクリーニング方法において使用されるGPR174は、GPR174としてのリゾホスファチジルセリン受容体活性を有するポリペプチドであれば特に限定されす、例えば、ヒト、マウス、ラットなどの哺乳類由来のGPR174が使用される。具体的な例としては、以下のポリペプチドが挙げられる。
GPR174 used in the screening method of the present invention is not particularly limited as long as it is a polypeptide having lysophosphatidylserine receptor activity as GPR174. For example, GPR174 derived from mammals such as humans, mice and rats is used. . Specific examples include the following polypeptides.
(a)配列番号22~24で表されるアミノ酸配列からなるポリペプチド;
(b)配列番号22~24で表されるアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド;
(c)配列番号22~24で表されるアミノ酸配列において1~10のアミノ酸が欠失、置換、付加、および/または挿入されたアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド;
(d)配列番号22~24で表されるアミノ酸配列との相同性が80%以上、好ましくは90%以上、より好ましくは95%以上であるアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド。 (A) a polypeptide comprising an amino acid sequence represented by SEQ ID NOs: 22 to 24;
(B) a polypeptide comprising the amino acid sequence represented by SEQ ID NOs: 22 to 24 and having lysophosphatidylserine receptor activity;
(C) a polypeptide comprising an amino acid sequence in which 1 to 10 amino acids are deleted, substituted, added and / or inserted in the amino acid sequence represented by SEQ ID NOs: 22 to 24 and having lysophosphatidylserine receptor activity ;
(D) an amino acid sequence having a homology with the amino acid sequences represented by SEQ ID NOs: 22 to 24 of 80% or more, preferably 90% or more, more preferably 95% or more, and having lysophosphatidylserine receptor activity Having a polypeptide.
(b)配列番号22~24で表されるアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド;
(c)配列番号22~24で表されるアミノ酸配列において1~10のアミノ酸が欠失、置換、付加、および/または挿入されたアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド;
(d)配列番号22~24で表されるアミノ酸配列との相同性が80%以上、好ましくは90%以上、より好ましくは95%以上であるアミノ酸配列を含み、かつリゾホスファチジルセリン受容体活性を有するポリペプチド。 (A) a polypeptide comprising an amino acid sequence represented by SEQ ID NOs: 22 to 24;
(B) a polypeptide comprising the amino acid sequence represented by SEQ ID NOs: 22 to 24 and having lysophosphatidylserine receptor activity;
(C) a polypeptide comprising an amino acid sequence in which 1 to 10 amino acids are deleted, substituted, added and / or inserted in the amino acid sequence represented by SEQ ID NOs: 22 to 24 and having lysophosphatidylserine receptor activity ;
(D) an amino acid sequence having a homology with the amino acid sequences represented by SEQ ID NOs: 22 to 24 of 80% or more, preferably 90% or more, more preferably 95% or more, and having lysophosphatidylserine receptor activity Having a polypeptide.
ここで相同性の数値は、当業者に公知の相同性検索プログラムを用いて算出される数値であってよく、例えば、AltschulらによるBLAST (Basic Local Alignment Search Tool) プログラム(たとえば、Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ., J. Mol. Biol., 215: p403-410 (1990), Altschyl SF, Madden TL, Schaffer AA, Zhang J, Miller W, Lipman DJ., Nucleic Acids Res. 25: p3389-3402 (1997))を利用し決定することができる。例えば、アミノ酸配列については、BLAST2 〔Nucleic Acids Res.、 25、 3389(1997); Genome Res.、 7、 649 (1997)〕においてデフォルトパラメータを用いて算出される数値などがあげられる。
Here, the numerical value of homology may be a numerical value calculated using a homology search program known to those skilled in the art. For example, the BLAST (Basic Local Alignment Search Tool) program (for example, Altschul SF, Gish by Altschul et al. W, Miller W, Myers EW, Lipman DJ., J. Mol. Biol., 215: p403-410 (1990), Altschyl SF, Madden TL, Schaffer AA, Zhang J, Miller W, Lipman DJ, Acids . 25: p3389-3402 (1997)). For example, as for the amino acid sequence, numerical values calculated using default parameters in BLAST2N [Nucleic Acids Res., 25, (3389 (1997); Genome な ど Res., 7, 649 (1997)] can be mentioned.
本発明において特定のアミノ酸配列の変異体や相同体を使用する場合には、変異体や相同体のアミノ酸配列をコードするDNAを用いて、当該変異体または相同体を調製することができる。このような変異体や相同体をコードするDNAは、化学合成、遺伝子工学的手法、突然変異誘発などの当業者に既知の任意の方法で作製することもできる。具体的には、配列番号13~18に記載の配列をコードするDNAを利用し、これらDNAに変異を導入することにより所望のDNAを取得することができる。例えば、DNAに対し、変異原となる薬剤と接触作用させる方法、紫外線を照射する方法、遺伝子工学的手法等を用いて行うことができる。遺伝子工学的手法の一つである部位特異的変異誘発法は特定の位置に特定の変異を導入できる手法であることから有用であり、モレキュラークローニング第2版、カレント・プロトコールズ・イン・モレキュラー・バイオロジー、Nucleic Acids Research, 10, 6487, 1982、Nucleic Acids Research, 12, 9441, 1984、Nucleic Acids Research, 13, 4431, 1985、Nucleic Acids Research, 13, 8749,1985、Proc. Natl. Acad. Sci. USA, 79, 6409, 1982、Proc. Natl. Acad. Sci.USA, 82, 488, 1985、Gene, 34, 315, 1985、Gene, 102, 67, 1991等に記載の方法に準じて行うことができる。
In the present invention, when a mutant or homologue of a specific amino acid sequence is used, the mutant or homologue can be prepared using DNA encoding the amino acid sequence of the mutant or homologue. DNA encoding such a mutant or homologue can also be prepared by any method known to those skilled in the art, such as chemical synthesis, genetic engineering techniques, and mutagenesis. Specifically, DNA encoding the sequences described in SEQ ID NOs: 13 to 18 is used, and desired DNA can be obtained by introducing mutations into these DNAs. For example, it can be carried out using a method of bringing DNA into contact with a drug that becomes a mutagen, a method of irradiating ultraviolet rays, a genetic engineering method, or the like. Site-directed mutagenesis, which is one of the genetic engineering methods, is useful because it can introduce a specific mutation at a specific position. Molecular cloning 2nd edition, Current Protocols in Molecular. Biology, Nucleic Acids Research, 10, 6487, 1982, Nucleic Acids Research, 12, 9441, 1984, Nucleic Acids Research, 13, 4431, 1985, Nucleic Acids Research, 13, 8749,1985, Proc. Na Sci.Acad. USA, 79, 6409, 1982, Proc. Natl. Acad. Sci. USA, 82, 488, 1985, Gene, 34, 315, 1985, Gene, 102, 67, 1991, etc. Can do.
本発明のスクリーニングにおいて、対照化合物としてリゾホスファチジルセリンまたはその類縁体を使用することができ、その例としては上記式(I)、(Ia)および(Ib)の化合物など、より具体的には後述の実施例に記載した化合物などが挙げられる。
In the screening of the present invention, lysophosphatidylserine or an analog thereof can be used as a control compound, examples of which include the compounds of the above formulas (I), (Ia) and (Ib), and more specifically And the compounds described in the examples.
本明細書において「C1-3アルキル」とは、炭素数1~3の直鎖状、分岐鎖状、環状のアルキル基を意味し、例えば、メチル、エチル、n-プロピル、i-プロピル、シクロプロピルが含まれる。
In the present specification, “C 1-3 alkyl” means a linear, branched, or cyclic alkyl group having 1 to 3 carbon atoms, such as methyl, ethyl, n-propyl, i-propyl, Cyclopropyl is included.
本明細書において「C1-6アルキル」とは、炭素数1~6の直鎖状、分岐鎖状、環状または部分的に環状のアルキル基を意味し、例えば、メチル、エチル、n-プロピル、i-プロピル、n-ブチル、s-ブチル、i-ブチル、t-ブチル、n-ペンチル、3-メチルブチル、2-メチルブチル、1-メチルブチル、1-エチルプロピル、n-ヘキシル、4-メチルペンチル、3-メチルペンチル、2-メチルペンチル、1-メチルペンチル、3-エチルブチル、および2-エチルブチル、シクロプロピル、シクロブチル、シクロペンチル、シクロヘキシル、およびシクロプロピルメチルなどが含まれ、例えば、C1-4アルキルおよびC1-3アルキルなども含まれる。
In the present specification, “C 1-6 alkyl” means a linear, branched, cyclic or partially cyclic alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl. I-propyl, n-butyl, s-butyl, i-butyl, t-butyl, n-pentyl, 3-methylbutyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, n-hexyl, 4-methylpentyl , 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-ethylbutyl, and 2-ethylbutyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, and the like, for example, C 1-4 alkyl And C 1-3 alkyl and the like are also included.
本明細書の化合物の定義において「C1-10アルキル」とは、炭素数1~10の直鎖状、分岐鎖状、環状または部分的に環状のアルキル基を意味し、例えば、C1-6アルキルとして既に例示したアルキル基のほか、C7H15、C8H17、C9H19、およびC10H21で表される、直鎖状、分岐鎖状、環状または部分的に環状ののアルキル基が含まれる。
The "C 1-10 alkyl" in the definition of the compounds herein, refers to a linear, branched, cyclic or partially cyclic alkyl group having 1 to 10 carbon atoms, e.g., C 1- In addition to the alkyl groups already exemplified as 6 alkyl, linear, branched, cyclic or partially cyclic represented by C 7 H 15 , C 8 H 17 , C 9 H 19 , and C 10 H 21 These alkyl groups are included.
本明細書において「C3-30アルキル」とは、炭素数7~30の直鎖状、分岐鎖状、環状または部分的に環状のアルキル基を意味し、例えば、C1-6アルキルとして既に例示したC3-6アルキル基のほか、C7H15、C8H17、C9H19、C10H21、C11H23、C12H25、C13H27、C14H29、C15H31、C16H33、C17H35、C18H37、C19H39、C20H41、C21H43、C22H45、C23H47、C24H49、C25H51、C26H53、C27H55、C28H57、C29H59、およびC30H61で表される、直鎖状または分岐鎖状、環状または部分的に環状のアルキル基が含まれる。
In the present specification, “C 3-30 alkyl” means a linear, branched, cyclic or partially cyclic alkyl group having 7 to 30 carbon atoms, such as C 1-6 alkyl. In addition to the exemplified C 3-6 alkyl group, C 7 H 15 , C 8 H 17 , C 9 H 19 , C 10 H 21 , C 11 H 23 , C 12 H 25 , C 13 H 27 , C 14 H 29 , C 15 H 31 , C 16 H 33 , C 17 H 35 , C 18 H 37 , C 19 H 39 , C 20 H 41 , C 21 H 43 , C 22 H 45 , C 23 H 47 , C 24 H 49 , C 25 H 51 , C 26 H 53 , C 27 H 55 , C 28 H 57 , C 29 H 59 , and C 30 H 61 , linear or branched, cyclic or partially cyclic Contains alkyl groups .
本明細書において「C1-6アルキルカルボニル」とは、アルキル部分として既に定義したC1-6アルキル基を有するアルキルカルボニル基を意味し、例えばメチルカルボニル(アセチル)、エチルカルボニル、tert-ブチルカルボニルの他、C1-4アルキルカルボニルなどが含まれる。
As used herein, “C 1-6 alkylcarbonyl” means an alkylcarbonyl group having a C 1-6 alkyl group already defined as the alkyl moiety, such as methylcarbonyl (acetyl), ethylcarbonyl, tert-butylcarbonyl. In addition, C 1-4 alkylcarbonyl and the like are included.
本明細書において「C1-6アルコキシ」とは、アルキル部分として既に定義した炭素数1~6のアルキル基を有するアルキルオキシ基を意味し、例えば、メトキシ、エトキシ、n-プロポキシ、i-プロポキシ、n-ブトキシ、s-ブトキシ、i-ブトキシ、t-ブトキシ、n-ペントキシ、3-メチルブトキシ、2-メチルブトキシ、1-メチルブトキシ、1-エチルプロポキシ、n-ヘキシルオキシ、4-メチルペントキシ、3-メチルペントキシ、2-メチルペントキシ、1-メチルペントキシ、3-エチルブトキシ、シクロペンチルオキシ、シクロヘキシルオキシ、シクロプロピルメチルオキシなどが含まれ、例えば、C1-4アルコキシおよびC1-3アルコキシなども含まれる。また、本明細書において「C1-4アルコキシ」には、例えばC1-3アルコキシなども含まれる。
In the present specification, “C 1-6 alkoxy” means an alkyloxy group having an alkyl group having 1 to 6 carbon atoms already defined as an alkyl moiety, and includes, for example, methoxy, ethoxy, n-propoxy, i-propoxy N-butoxy, s-butoxy, i-butoxy, t-butoxy, n-pentoxy, 3-methylbutoxy, 2-methylbutoxy, 1-methylbutoxy, 1-ethylpropoxy, n-hexyloxy, 4-methylpen Toxic, 3-methylpentoxy, 2-methylpentoxy, 1-methylpentoxy, 3-ethylbutoxy, cyclopentyloxy, cyclohexyloxy, cyclopropylmethyloxy, etc. are included, for example, C 1-4 alkoxy and C 1 Also included are -3 alkoxy and the like. In this specification, “C 1-4 alkoxy” includes, for example, C 1-3 alkoxy and the like.
本明細書において「C1-6アルコキシカルボニル」とは、アルコキシ部分として既に定義したC1-6アルコキシ基を有するアルコキシカルボニル基を意味し、例えばメトキシカルボニル、エトキシカルボニル、tert-ブトキシカルボニルの他、C1-3アルコキシカルボニルなどが含まれる。
As used herein, “C 1-6 alkoxycarbonyl” means an alkoxycarbonyl group having a C 1-6 alkoxy group already defined as an alkoxy moiety, such as methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, C 1-3 alkoxycarbonyl and the like are included.
本明細書における「C1-6アルコキシC1-6アルキル」には、例えばメトキシメチル、エトキシメチル、2-メトキシエチル、1-メトキシエチルなどが含まれる。
The “C 1-6 alkoxy C 1-6 alkyl” in the present specification includes, for example, methoxymethyl, ethoxymethyl, 2-methoxyethyl, 1-methoxyethyl and the like.
本明細書におけるベンジルは、式:C6H5CH2-で表される基を意味する。本明細書におけるベンジルオキシは、式:C6H5CH2O-で表される基を意味する。本明細書におけるベンジルオキシカルボニルは、式:C6H5CH2OC(=O)-で表される基を意味する。
In the present specification, benzyl means a group represented by the formula: C 6 H 5 CH 2 —. In the present specification, benzyloxy means a group represented by the formula: C 6 H 5 CH 2 O—. In the present specification, benzyloxycarbonyl means a group represented by the formula: C 6 H 5 CH 2 OC (═O) —.
本明細書において「5-6員へテロアリール」とは、酸素原子、窒素原子および硫黄原子から選択される1以上のヘテロ原子を含む、環原子が5~6の単環の芳香族ヘテロ環基を意味する。具体例としては、ピロリル、イミダゾリル、ピラゾリル、トリアゾリル、ピリジル、ピリミジル、ピリダジニル、フリル、チエニル、オキサゾリル、オキサジアゾリル、チアゾリル、チアジアゾリルなどが含まれる。
As used herein, “5-6-membered heteroaryl” refers to a monocyclic aromatic heterocyclic group having 5 to 6 ring atoms, including one or more heteroatoms selected from an oxygen atom, a nitrogen atom and a sulfur atom Means. Specific examples include pyrrolyl, imidazolyl, pyrazolyl, triazolyl, pyridyl, pyrimidyl, pyridazinyl, furyl, thienyl, oxazolyl, oxadiazolyl, thiazolyl, thiadiazolyl and the like.
本明細書において「フェニレン」とは、ベンゼンが2カ所で置換された2価の基を意味し、1,2-置換、1,3-置換、および1,4-置換の何れであってもよい。
In the present specification, “phenylene” means a divalent group in which benzene is substituted at two positions, and may be any of 1,2-substituted, 1,3-substituted, and 1,4-substituted. Good.
本明細書において「5-6員へテロアリーレン」とは、酸素原子、窒素原子および硫黄原子から選択される1以上のヘテロ原子を含む、環原子が5~6の単環の芳香族ヘテロ環が2カ所で置換された2価の基を意味する。当該基を構成するヘテロ環の具体例としては、ピロール、イミダゾール、ピラゾール、トリアゾール、ピリジン、ピリミジン、ピリダジニン、フラン、チオフェン、オキサゾール、オキサジアゾール、チアゾール、チアジアゾールなどが挙げられる。
As used herein, “5-6-membered heteroarylene” refers to a monocyclic aromatic heterocycle having 5 to 6 ring atoms, including one or more heteroatoms selected from an oxygen atom, a nitrogen atom, and a sulfur atom Means a divalent group substituted at two positions. Specific examples of the heterocyclic ring constituting the group include pyrrole, imidazole, pyrazole, triazole, pyridine, pyrimidine, pyridazine, furan, thiophene, oxazole, oxadiazole, thiazole, thiadiazole and the like.
本明細書において、アルキルに1以上のフェニレンまたは5-6員ヘテロアリーレンが挿入されている場合、フェニレンおよび5-6員ヘテロアリーレンの挿入場所、ならびにフェニレンおよび5-6員ヘテロアリーレンの置換位置は特に限定されない。挿入されるフェニレンおよび5-6員ヘテロアリーレンの数は、例えば1~5、好ましくは1~3である。
In the present specification, when one or more phenylene or 5-6 membered heteroarylene is inserted into alkyl, the insertion position of phenylene and 5-6 membered heteroarylene, and the substitution position of phenylene and 5-6 membered heteroarylene are There is no particular limitation. The number of inserted phenylene and 5-6 membered heteroarylene is, for example, 1 to 5, preferably 1 to 3.
本明細書において、アルキルの1以上の-CH2-が-O-と置き換えられている場合、置き換えられる-CH2-の位置は置き換え可能であれば特に限定されない。また置き換えられた-O-と-CH2-の間に、または-O-と-O-の間にフェニレンまたは5-6員ヘテロアリーレンが挿入されていてもよい。置き換えられる-O-の数は、例えば1~10、好ましくは1~5、より好ましくは1~3である。
In this specification, when one or more —CH 2 — of alkyl is replaced with —O—, the position of —CH 2 — to be replaced is not particularly limited as long as it can be replaced. Further, phenylene or a 5-6 membered heteroarylene may be inserted between the substituted —O— and —CH 2 — or between —O— and —O—. The number of —O— to be replaced is, for example, 1 to 10, preferably 1 to 5, and more preferably 1 to 3.
本明細書において、炭素原子間の1以上の単結合が、二重結合または三重結合に置き換えられている場合、置き換えられる単結合の位置は置き換え可能であれば特に限定されない。二重結合はシス配置であってもトランス配置であってもよい。置き換えられる二重結合および三重結合の数は、例えば1~10、好ましくは1~5、より好ましくは1~3である。
In the present specification, when one or more single bonds between carbon atoms are replaced with double bonds or triple bonds, the position of the replaced single bond is not particularly limited as long as it can be replaced. The double bond may be a cis configuration or a trans configuration. The number of double bonds and triple bonds replaced is, for example, 1 to 10, preferably 1 to 5, and more preferably 1 to 3.
上記式(I)、(Ia)、および(Ib)で表される化合物に関する本発明には、互変異性体、幾何異性体、光学異性体などの各種の立体異性体、およびそれらの混合物が含まれる。
In the present invention relating to the compounds represented by the above formulas (I), (Ia), and (Ib), various stereoisomers such as tautomers, geometric isomers, optical isomers, and mixtures thereof are included. included.
本発明の化合物の「医薬として許容な塩」とは、医薬品として使用可能な塩であれば特に限定されない。本発明化合物が塩基と形成する塩としては、ナトリウム、カリウム、マグネシウム、カルシウム、アルミニウムなどの無機塩基との塩;メチルアミン、エチルアミン、エタノールアミン等の有機塩基との塩などが挙げられる。当該塩は、酸付加塩であってもよく、かかる塩としては、具体的には、塩酸、臭化水素酸、ヨウ化水素酸、硫酸、硝酸、リン酸等の鉱酸;および、ギ酸、酢酸、プロピオン酸、シュウ酸、マロン酸、コハク酸、フマル酸、マレイン酸、乳酸、リンゴ酸、酒石酸、クエン酸、メタンスルホン酸、エタンスルホン酸などの有機酸酸との酸付加塩が挙げられる。
The “pharmaceutically acceptable salt” of the compound of the present invention is not particularly limited as long as it is a salt that can be used as a pharmaceutical product. Examples of the salt formed by the compound of the present invention with a base include salts with inorganic bases such as sodium, potassium, magnesium, calcium and aluminum; salts with organic bases such as methylamine, ethylamine and ethanolamine. The salt may be an acid addition salt. Specific examples of the salt include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid and other mineral acids; and formic acid, Examples include acid addition salts with organic acid acids such as acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, and ethanesulfonic acid. .
更に、本発明化合物には、水和物、各種溶媒和物や結晶多形等も含まれる。
Furthermore, the compounds of the present invention also include hydrates, various solvates and crystal polymorphs.
本発明の化合物に含まれる原子(例えば、水素原子、炭素原子、酸素原子、窒素原子、硫黄原子およびリン原子など)は、それぞれの天然に最も多く存在する同位体以外の同位体原子であってもよく、当該同位体原子は放射性導体原子であってもよい。すなわち、本発明の1つの側面によれば、同位体原子で標識化された本明細書で既に定義された式(I)、(Ia)または(Ib)の化合物、またはその塩が提供される。ここで、同位体原子による標識化は、例えば、放射性同位体による標識化(3H、14C、32Pなど)であってもよく、化合物の調製の容易さの側面からは、3Hによる標識化が好ましい。本発明の3H標識化化合物は、例えば、3H標識化された脂肪酸またはその誘導体を使用することにより合成することができる。
The atoms (for example, hydrogen atom, carbon atom, oxygen atom, nitrogen atom, sulfur atom and phosphorus atom) contained in the compound of the present invention are isotope atoms other than the most naturally occurring isotopes. The isotope atom may be a radioactive conductor atom. That is, according to one aspect of the present invention there is provided a compound of formula (I), (Ia) or (Ib) as defined herein, or a salt thereof, labeled with an isotope atom. . Here, labeling with an isotope atom may be, for example, labeling with a radioisotope ( 3 H, 14 C, 32 P, etc.). From the aspect of ease of preparation of the compound, 3 H Labeling is preferred. The 3 H-labeled compound of the present invention can be synthesized, for example, by using a 3 H-labeled fatty acid or a derivative thereof.
本発明の1つの態様において、式(I)、(Ia)および(Ib)の化合物は、プロドラッグとして投与され、生体内において活性化合物に変換される。例えば式(I)のR5、(Ia)のR15および(Ib)のR25はリン酸エステルを形成する基であってもよい。その具体例としては、例えばJournal of Medicinal Chemistry, 2008, 51(8), 2337に記載の基などが挙げられ、C1-6アルキル(例えば、tert-ブチル)、C1-6アルコキシC1-6アルキル(例えば、メトキシメチル)、C1-6アルキルカルボニルオキシC1-6アルキル(例えば、ピバロイルオキシメチル)、C1-6アルコキシカルボニルオキシC1-6アルキル(例えば、イソプロポキシカルボニルオキシメチル)、置換されていてもよいフェニル(例えば、C1-3アルコキシフェニル)、置換されていてもよいベンジル(例えば、C1-6アルコキシ、ヒドロキシ、C1-6アルキルカルボニルオキシ、ニトロ、およびハロゲン原子から選択される1~3の基により置換されていてもよいベンジル)、フタリジル(例えば、C1-6アルコキシから選択される1~4の基により置換されていてもよいイソベンゾフラノン-3-イル)、ジオキソレノンイルメチル(例えば、ジオキソレノン環5位がC1-6アルコキシまたはフェニルから選択される基により置換されていてもよいジオキソレノン-4-イルメチル)、またはフリルメチル(例えば、フラン環5位がニトロにより置換されていてもよい2-フリルメチルなどが含まれる。
In one embodiment of the invention, the compounds of formula (I), (Ia) and (Ib) are administered as prodrugs and are converted in vivo into active compounds. For example, R 5 of formula (I), R 15 of (Ia) and R 25 of (Ib) may be a group forming a phosphate ester. Specific examples thereof include groups described in Journal of Medicinal Chemistry, 2008, 51 (8), 2337, and the like. C 1-6 alkyl (eg, tert-butyl), C 1-6 alkoxy C 1- 6 alkyl (eg methoxymethyl), C 1-6 alkylcarbonyloxy C 1-6 alkyl (eg pivaloyloxymethyl), C 1-6 alkoxycarbonyloxy C 1-6 alkyl (eg isopropoxycarbonyloxy) Methyl), optionally substituted phenyl (eg, C 1-3 alkoxyphenyl), optionally substituted benzyl (eg, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro, and benzyl which may be substituted by a group of 1 to 3 selected from a halogen atom), phthalidyl (e.g., C 1-6 Good -isobenzofuranone-3-yl optionally substituted by a group having 1 to 4 selected from alkoxy), di-oxo Lennon yl methyl (e.g., Jiokisorenon ring 5-position is selected from C 1-6 alkoxy or phenyl Dioxolenon-4-ylmethyl which may be substituted by a group, or furylmethyl (for example, 2-furylmethyl which may be substituted at the 5-position of the furan ring by nitro).
式(I)、(Ia)および(Ib)の化合物は、例えば、以下のスキームに示す工程により合成することができる。
Compounds of formula (I), (Ia) and (Ib) can be synthesized, for example, by the steps shown in the following scheme.
[式中、A、n、X、Y、R5、およびR6は、本明細書において既に定義したとおりであり;R1aおよびR2aは、それぞれ独立にC1-6アルキルから選択される]
Wherein A, n, X, Y, R 5 , and R 6 are as previously defined herein; R 1a and R 2a are each independently selected from C 1-6 alkyl ]
式(I)の化合物は、化合物1-1と化合物1-2を縮合した後に酸化することにより調製することができる。第1段階の縮合反応は例えば塩化メチレン、テトラヒドロフラン、N,N’-ジメチルホルムアミド、トルエン、ジエチルエーテル、1,4-ジオキサンなどの適当な溶媒中で、適当な反応促進剤(例えば1H-テトラゾールなど)の存在下で行うことができる。この工程は、特に限定はされないが、例えば0~70℃、好ましくは15~30℃の反応温度、および例えば10分~2日、好ましくは1~2時間の反応時間で行うことができる。また、本工程においては化合物1-1と化合物1-2を適当な溶媒に溶解し、混合した後にトルエンやベンゼンなどの溶媒により共沸処理を施すことが望ましい。
The compound of the formula (I) can be prepared by condensing the compound 1-1 and the compound 1-2 and then oxidizing. The first stage condensation reaction is carried out in a suitable solvent such as methylene chloride, tetrahydrofuran, N, N′-dimethylformamide, toluene, diethyl ether, 1,4-dioxane, etc. (for example, 1H-tetrazole, etc.) ) In the presence of This step is not particularly limited, but can be performed, for example, at a reaction temperature of 0 to 70 ° C., preferably 15 to 30 ° C., and a reaction time of, for example, 10 minutes to 2 days, preferably 1 to 2 hours. In this step, it is desirable that compound 1-1 and compound 1-2 are dissolved in a suitable solvent, mixed and then subjected to azeotropic treatment with a solvent such as toluene or benzene.
第2段階の酸化工程は、例えば塩化メチレン、テトラヒドロフラン、N,N’-ジメチルホルムアミド、トルエン、ジエチルエーテル、1,4-ジオキサンなどの適当な溶媒中で、適当な酸化剤(例えばt-ブチルヒドロペルオキシド、メタクロロ過安息香酸、ヨウ素-ピリジン-水等)を使用して行うことができる。この工程は、特に限定はされないが、例えば0~70℃、好ましくは15~30℃の反応温度、および例えば5分~1日、好ましくは30分~1時間の反応時間で行うことができる。第2段階の酸化反応は、第1段階の縮合反応を後処理して得られた生成物に対して行うこともできるが、第1段階の後処理を行うことなしにワンポット反応として行ってもよい。
The second stage oxidation step is carried out in a suitable solvent such as, for example, tert-butylhydrochloride in a suitable solvent such as methylene chloride, tetrahydrofuran, N, N′-dimethylformamide, toluene, diethyl ether, 1,4-dioxane. Peroxide, metachloroperbenzoic acid, iodine-pyridine-water, etc.). This step is not particularly limited, but can be performed, for example, at a reaction temperature of 0 to 70 ° C., preferably 15 to 30 ° C., and a reaction time of, for example, 5 minutes to 1 day, preferably 30 minutes to 1 hour. The oxidation reaction in the second stage can be performed on the product obtained by post-treating the condensation reaction in the first stage, but can be performed as a one-pot reaction without performing the post-treatment in the first stage. Good.
得られる化合物1-3が保護基を有する場合は、適切な脱保護条件に付すことにより、および/または、さらに置換基変換を行うことにより、目的の式(I)、(Ia)および(Ib)の化合物を得ることができる。
When the obtained compound 1-3 has a protecting group, it can be subjected to appropriate deprotection conditions and / or by further substituent substitution to obtain the desired formula (I), (Ia) and (Ib ) Can be obtained.
[式中、A、n、X、R5、およびR6は、本明細書において既に定義したとおりであり、Yは-OC(=O)-である]
[Wherein A, n, X, R 5 , and R 6 are as defined herein, and Y is —OC (═O) —]
式(I)の化合物は、化合物2-1を化合物2-2の酸クロリドでアシル化することにより調製することができる。当該反応は例えば塩化メチレン、テトラヒドロフラン、N,N’-ジメチルホルムアミド、トルエン、ジエチルエーテル、1,4-ジオキサンなどの適当な溶媒中で、適当な塩基(例えば4-ジメチルアミノピリジンなど)の存在下または塩基の非存在下で行うことができる。この工程は、特に限定はされないが、例えば0~70℃、好ましくは15~30℃の反応温度、および例えば10分~2日の反応時間で行うことができる。アシル化は、化合物2-2に対応するカルボン酸と適当な縮合剤(例えば、1-エチル-3-(3-ジメチルアミノプロピル)カルボジイミド塩酸塩など)を使用して行うこともできる。
The compound of formula (I) can be prepared by acylating compound 2-1 with an acid chloride of compound 2-2. The reaction is carried out in a suitable solvent such as methylene chloride, tetrahydrofuran, N, N′-dimethylformamide, toluene, diethyl ether, 1,4-dioxane and the like in the presence of a suitable base (for example, 4-dimethylaminopyridine). Alternatively, it can be performed in the absence of a base. This step is not particularly limited, but can be performed at a reaction temperature of, for example, 0 to 70 ° C., preferably 15 to 30 ° C., and a reaction time of, for example, 10 minutes to 2 days. The acylation can also be carried out using a carboxylic acid corresponding to compound 2-2 and a suitable condensing agent (for example, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride).
得られる化合物2-3が保護基を有する場合は、適切な脱保護条件に付すことにより、および/または、さらに置換基変換を行うことにより、目的の式(I)、(Ia)および(Ib)の化合物を得ることができる。
When the obtained compound 2-3 has a protecting group, it can be subjected to appropriate deprotection conditions and / or by further substituent substitution to obtain the desired formula (I), (Ia) and (Ib). ) Can be obtained.
本発明の医薬組成物は、種々の剤形、例えば、経口投与のためには、錠剤、カプセル剤、散剤、顆粒剤、丸剤、液剤、乳剤、懸濁液、溶液剤、酒精剤、シロップ剤、エキス剤、エリキシル剤とすることができ、非経口剤としては、例えば、皮下注射剤、静脈内注射剤、筋肉内注射剤、腹腔内注射剤などの注射剤;経皮投与または貼付剤、軟膏またはローション;口腔内投与のための舌下剤、口腔貼付剤;ならびに経鼻投与のためのエアゾール剤とすることができるが、これらには限定されない。これらの製剤は、製剤工程において通常用いられる公知の方法により製造することができる。式(I)、(Ia)および(Ib)の化合物は、好ましくは非経口剤として投与される。
The pharmaceutical composition of the present invention can be used in various dosage forms such as tablets, capsules, powders, granules, pills, solutions, emulsions, suspensions, solutions, spirits, syrups for oral administration. And parenteral preparations include, for example, injections such as subcutaneous injections, intravenous injections, intramuscular injections, intraperitoneal injections; transdermal administration or patches , Ointments or lotions; sublingual and buccal patches for buccal administration; and aerosols for nasal administration, but not limited thereto. These preparations can be produced by known methods usually used in the preparation process. The compounds of formula (I), (Ia) and (Ib) are preferably administered as parenteral agents.
当該医薬組成物は、一般に用いられる各種成分を含みうるものであり、例えば、1種以上の薬学的に許容され得る賦形剤、崩壊剤、希釈剤、滑沢剤、着香剤、着色剤、甘味剤、矯味剤、懸濁化剤、湿潤剤、乳化剤、分散剤、補助剤、防腐剤、緩衝剤、結合剤、安定剤、コーティング剤等を含みうる。また本発明の医薬組成物は、持続性または徐放性剤形であってもよい。
The pharmaceutical composition may contain various commonly used components, such as one or more pharmaceutically acceptable excipients, disintegrants, diluents, lubricants, flavoring agents, and coloring agents. , Sweeteners, flavoring agents, suspending agents, wetting agents, emulsifying agents, dispersing agents, adjuvants, preservatives, buffering agents, binders, stabilizers, coating agents and the like. The pharmaceutical composition of the present invention may be in a sustained or sustained release dosage form.
本発明の治療剤、予防剤、または医薬組成物の投与量は、投与経路、患者の体型、年齢、体調、疾患の度合い、発症後の経過時間等により、適宜選択することができ、本発明の医薬組成物は、治療有効量および/または予防有効量の上記式(I)、(Ia)、または(Ib)の化合物を含むことができる。本発明において上記式(I)、式(Ia)または式(Ib)の化合物は、一般に1~10000mg/日/成人の用量で使用されうる。当該医薬組成物の投与は、単回投与または複数回投与であってもよく、例えば免疫抑制剤(シクロスポリン、タクロリムス、シロリムス、、メトトレキサート、アザチオプリンなど)、ステロイド系抗炎症薬(ヒドロコルチゾン、プレドニゾロン、デキサメタゾンなど)、非ステロイド系抗炎症薬(ロキソプロフェンナトリウム、インドメタシン、ジクロフェナクナトリウムなど)、または抗体医薬(インフリキシマブ、アダリムマブ、トシリズマブ、セルトリズマブペゴル、エタネルセプトなど)などの他の薬剤と組み合わせて使用することもできる。
The dosage of the therapeutic agent, prophylactic agent, or pharmaceutical composition of the present invention can be appropriately selected depending on the administration route, the patient's body shape, age, physical condition, degree of disease, elapsed time after onset, etc. The pharmaceutical composition can comprise a therapeutically effective amount and / or a prophylactically effective amount of a compound of formula (I), (Ia), or (Ib) above. In the present invention, the compound of the above formula (I), formula (Ia) or formula (Ib) can be generally used at a dose of 1 to 10000 mg / day / adult. The pharmaceutical composition may be administered in a single dose or multiple doses, such as immunosuppressants (cyclosporine, tacrolimus, sirolimus, methotrexate, azathioprine, etc.), steroidal anti-inflammatory drugs (hydrocortisone, prednisolone, dexamethasone) Etc.), non-steroidal anti-inflammatory drugs (such as loxoprofen sodium, indomethacin, diclofenac sodium), or antibody drugs (such as infliximab, adalimumab, tocilizumab, sertolizumab pegol, etanercept) You can also.
本発明の治療剤または予防剤は、必要に応じ、従来公知の着色剤、保存剤、香料、風味剤、コーティング剤、抗酸化剤、ビタミン、アミノ酸、ペプチド、タンパク質、およびミネラル分(鉄、亜鉛、マグネシム、ヨードなど)などの成分を含有していてもよい。本発明の治療剤または予防剤は、医薬組成物、機能性食品、健康食品、飲料、サプリメントなどに適した形態、例えば顆粒剤(ドライシロップを含む)、カプセル剤(軟カプセル剤、硬カプセル剤)、錠剤(チュアブル剤などを含む)、散剤(粉末剤)、丸剤などの各種の固形製剤、または内服用液剤(液剤、懸濁剤、シロップ剤を含む)などの液状製剤などの形態で調製してもよい。また、本発明の治療剤または予防剤は、そのまま、医薬組成物、機能性食品、健康食品、サプリメントなどとして使用することもできる。
The therapeutic agent or prophylactic agent of the present invention may contain a conventionally known coloring agent, preservative, flavoring, flavoring agent, coating agent, antioxidant, vitamin, amino acid, peptide, protein, and mineral content (iron, zinc, if necessary). , Magnesium, iodine, etc.). The therapeutic agent or prophylactic agent of the present invention is in a form suitable for pharmaceutical compositions, functional foods, health foods, beverages, supplements, etc., such as granules (including dry syrup), capsules (soft capsules, hard capsules). , Prepared in the form of various solid preparations such as tablets (including chewables), powders (powder), pills, or liquid preparations for internal use (including liquids, suspensions, syrups) May be. In addition, the therapeutic agent or prophylactic agent of the present invention can be used as it is as a pharmaceutical composition, functional food, health food, supplement or the like.
製剤化のための添加物としては、例えば、賦形剤、滑沢剤、結合剤、崩壊剤、流動化剤、分散剤、湿潤剤、防腐剤、粘稠剤、pH調整剤、着色剤、矯味矯臭剤、界面活性剤、溶解補助剤が挙げられる。また、液剤の形態にする場合は、ペクチン、キサンタンガム、グアガムなどの増粘剤を配合することができる。また、コーティング剤を用いてコーティング錠剤にしたり、ペースト状の膠剤とすることもできる。さらに、他の形態に調製する場合であっても、従来の方法に従えばよい。
As additives for formulation, for example, excipients, lubricants, binders, disintegrants, fluidizers, dispersants, wetting agents, preservatives, thickeners, pH adjusters, colorants, Examples include flavoring agents, surfactants, and solubilizing agents. Moreover, when making it into the form of a liquid agent, thickeners, such as pectin, xanthan gum, and guar gum, can be mix | blended. Moreover, it can also be set as a coating tablet using a coating agent, or it can also be set as a paste-form glue. Furthermore, even if it is a case where it prepares in another form, what is necessary is just to follow the conventional method.
以下、実施例を示すことにより本発明をさらに詳細に説明するが、本発明はこれらの実施例に限定されるものではない。
Hereinafter, the present invention will be described in more detail by showing examples, but the present invention is not limited to these examples.
試薬およびデータ測定
試薬はSigma-Aldrich Chemical Co.、東京化成工業、和光純薬、関東化学から購入したものをさらに精製することなく使用した。1H-および13C-NMRは、BRUKER AVANCE400スペクトロメーター(400MHz)を使用して測定し、ケミカルシフトは重クロロホルム(7.26ppm(1H-NMR)、77.00ppm(13C-NMR))に対するppmで表示した。31P-NMRケミカルシフトは水中のリン酸(85%w/w、0.00ppm)に対するppmで表示した。質量分析はBRUKER microTOF-05スペクトロメーター(ESI-TOF)またはa SHIMADZU AXIMA-TOF (MALDI-TOF)のポジティブおよびネガティブイオンモードまたはで測定した。カラムクロマトグラフィーに使用するシリカゲルは関東化学から購入した。元素分析はYanaco MT-6 CHN CORDERスペクトロメーターを使用して行った。 Reagents and data measurement reagents are available from Sigma-Aldrich Chemical Co. Those purchased from Tokyo Chemical Industry, Wako Pure Chemicals, and Kanto Chemical were used without further purification. 1 H- and 13 C-NMR were measured using aBRUKER AVANCE 400 spectrometer (400 MHz), and the chemical shift was deuterated chloroform (7.26 ppm ( 1 H-NMR), 77.00 ppm ( 13 C-NMR)). In ppm. 31 P-NMR chemical shifts are expressed in ppm relative to phosphoric acid in water (85% w / w, 0.00 ppm). Mass spectrometry was measured in the BRUKER microTOF-05 spectrometer (ESI-TOF) or a SHIMADZU AXIMA-TOF (MALDI-TOF) positive and negative ion mode. Silica gel used for column chromatography was purchased from Kanto Chemical. Elemental analysis was performed using a Yanaco MT-6 CHN CORDER spectrometer.
試薬はSigma-Aldrich Chemical Co.、東京化成工業、和光純薬、関東化学から購入したものをさらに精製することなく使用した。1H-および13C-NMRは、BRUKER AVANCE400スペクトロメーター(400MHz)を使用して測定し、ケミカルシフトは重クロロホルム(7.26ppm(1H-NMR)、77.00ppm(13C-NMR))に対するppmで表示した。31P-NMRケミカルシフトは水中のリン酸(85%w/w、0.00ppm)に対するppmで表示した。質量分析はBRUKER microTOF-05スペクトロメーター(ESI-TOF)またはa SHIMADZU AXIMA-TOF (MALDI-TOF)のポジティブおよびネガティブイオンモードまたはで測定した。カラムクロマトグラフィーに使用するシリカゲルは関東化学から購入した。元素分析はYanaco MT-6 CHN CORDERスペクトロメーターを使用して行った。 Reagents and data measurement reagents are available from Sigma-Aldrich Chemical Co. Those purchased from Tokyo Chemical Industry, Wako Pure Chemicals, and Kanto Chemical were used without further purification. 1 H- and 13 C-NMR were measured using a
[実施例1](2S)-2-アミノ-3-[(3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 1] (2S) -2-Amino-3-[(3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
工程1:2-tert-ブチル-1,3-ジイソプロピル-イソウレアの調製
Step 1: Preparation of 2-tert-butyl-1,3-diisopropyl-isourea
1,3-ジイソプロピルカルボジイミド(6.325g、50.1mmol)およびCuCl(50.8mg、0.513mmol)を無水tBuOH(4.556g、59.9mmol)に溶解させ、アルゴン雰囲気下、室温で3日間攪拌した。反応混合物の一部を少量とり、1H-NMRにより反応の完了を確認した。無水CH2Cl2(25ml)およびポリビニルピロリドン(1.004g)を反応混合物に加え、15分間攪拌し、セライト(登録商標)で濾過した。濾液を濃縮し、表題の化合物を得た(8.108g、40.47mmol、81%、緑色油状物)。得られた化合物は更に精製することなくエステル化試薬として使用した。
1,3-Diisopropylcarbodiimide (6.325 g, 50.1 mmol) and CuCl (50.8 mg, 0.513 mmol) were dissolved in anhydrous tBuOH (4.556 g, 59.9 mmol), and at room temperature under argon atmosphere for 3 days Stir. A small portion of the reaction mixture was taken and the completion of the reaction was confirmed by 1 H-NMR. Anhydrous CH 2 Cl 2 (25 ml) and polyvinylpyrrolidone (1.004 g) were added to the reaction mixture, stirred for 15 minutes and filtered through Celite®. The filtrate was concentrated to give the title compound (8.18 g, 40.47 mmol, 81%, green oil). The obtained compound was used as an esterification reagent without further purification.
1H-NMR(CDCl3):δ=3.760-3.609 (1H, m), 3.230-3.094 (1H, m), 1.456-1.363 (9H, m), 1.070 (6H, d, J=6.44 Hz), 1.036 (6H, d, J=6.16 Hz)。
1 H-NMR (CDCl 3 ): δ = 3.760-3.609 (1H, m), 3.230-3.094 (1H, m), 1.456-1.363 (9H, m), 1.070 (6H, d, J = 6.44 Hz), 1.036 (6H, d, J = 6.16 Hz).
工程2:N-tert-ブトキシカルボニルセリンtert-ブチルエステルの調製
Step 2: Preparation of N-tert-butoxycarbonylserine tert-butyl ester
1N NaOH水溶液(1N、10ml)、H2O(10ml)およびジオキサン(20ml)の混合液にL-セリン(1.025g、9.756mmol)を溶解させ、0℃で撹拌した。当該溶液にジ-tert-ブチル ジカーボネート(3.192g、14.63mmol)を0℃で加え、そのまま10分攪拌し、その後室温で24時間攪拌した。5%KHSO4水溶液を反応混合物に加え、pHを3にし、AcOEt(150ml×3)で抽出した。有機層を合わせて、食塩水で洗浄し、Na2SO4で乾燥させた。溶媒を留去して、N-Bocセリンを粗生成物として得た(2.228g、無色油状物)。N-Bocセリンの粗生成物(2.228g)を無水CH2Cl2(150ml)に溶解させ、2-tert-ブチル-1,3-ジイソプロピル-イソウレア(7.112g)を加え、反応溶液を室温で18時間攪拌した。反応混合物にヘキサン(150ml)を加え、10分攪拌した。該混合物をセライト(登録商標)で濾過し、濃縮した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=4:1)により精製し、表題の化合物を得た(1.677g、6.418mmol、66%、白色固体)。
1H-NMR(CDCl3):δ=5.407 (1H, m), 4.255 (1H, m), 3.897 (2H, m), 2.333 (1H, m), 1.483 (9H, s), 1.452 (9H, s)。
HRMS (ESI, [M+Na]+): 計算値 C12H23NNaO5 +: 284.1468;実測値:284.1469。
融点 76.9-77.7 ℃。 L-serine (1.025 g, 9.756 mmol) was dissolved in a mixture of 1N NaOH aqueous solution (1N, 10 ml), H 2 O (10 ml) and dioxane (20 ml), and the mixture was stirred at 0 ° C. Di-tert-butyl dicarbonate (3.192 g, 14.63 mmol) was added to the solution at 0 ° C., and the mixture was stirred as it was for 10 minutes and then at room temperature for 24 hours. 5% KHSO 4 aqueous solution was added to the reaction mixture to bring the pH to 3 and extracted with AcOEt (150 ml × 3). The organic layers were combined, washed with brine and dried over Na 2 SO 4 . The solvent was distilled off to give N-Boc serine as a crude product (2.228 g, colorless oil). The crude product of N-Boc serine (2.228 g) was dissolved in anhydrous CH 2 Cl 2 (150 ml), 2-tert-butyl-1,3-diisopropyl-isourea (7.112 g) was added, and the reaction solution was Stir at room temperature for 18 hours. Hexane (150 ml) was added to the reaction mixture and stirred for 10 minutes. The mixture was filtered through Celite® and concentrated. The residue was purified by column chromatography (hexane: AcOEt = 4: 1) to give the title compound (1.677 g, 6.418 mmol, 66%, white solid).
1 H-NMR (CDCl 3 ): δ = 5.407 (1H, m), 4.255 (1H, m), 3.897 (2H, m), 2.333 (1H, m), 1.483 (9H, s), 1.452 (9H, s).
HRMS (ESI, [M + Na] + ): calculated C 12 H 23 NNaO 5 + : 284.1468; found: 284.1469.
Melting point 76.9-77.7 ° C.
1H-NMR(CDCl3):δ=5.407 (1H, m), 4.255 (1H, m), 3.897 (2H, m), 2.333 (1H, m), 1.483 (9H, s), 1.452 (9H, s)。
HRMS (ESI, [M+Na]+): 計算値 C12H23NNaO5 +: 284.1468;実測値:284.1469。
融点 76.9-77.7 ℃。 L-serine (1.025 g, 9.756 mmol) was dissolved in a mixture of 1N NaOH aqueous solution (1N, 10 ml), H 2 O (10 ml) and dioxane (20 ml), and the mixture was stirred at 0 ° C. Di-tert-butyl dicarbonate (3.192 g, 14.63 mmol) was added to the solution at 0 ° C., and the mixture was stirred as it was for 10 minutes and then at room temperature for 24 hours. 5% KHSO 4 aqueous solution was added to the reaction mixture to bring the pH to 3 and extracted with AcOEt (150 ml × 3). The organic layers were combined, washed with brine and dried over Na 2 SO 4 . The solvent was distilled off to give N-Boc serine as a crude product (2.228 g, colorless oil). The crude product of N-Boc serine (2.228 g) was dissolved in anhydrous CH 2 Cl 2 (150 ml), 2-tert-butyl-1,3-diisopropyl-isourea (7.112 g) was added, and the reaction solution was Stir at room temperature for 18 hours. Hexane (150 ml) was added to the reaction mixture and stirred for 10 minutes. The mixture was filtered through Celite® and concentrated. The residue was purified by column chromatography (hexane: AcOEt = 4: 1) to give the title compound (1.677 g, 6.418 mmol, 66%, white solid).
1 H-NMR (CDCl 3 ): δ = 5.407 (1H, m), 4.255 (1H, m), 3.897 (2H, m), 2.333 (1H, m), 1.483 (9H, s), 1.452 (9H, s).
HRMS (ESI, [M + Na] + ): calculated C 12 H 23 NNaO 5 + : 284.1468; found: 284.1469.
Melting point 76.9-77.7 ° C.
工程3:(2S)-3-[(ジイソプロピルアミノ)-tert-ブトキシホスフィノオキシ]-2-(tert-ブトキシカルボニルアミノ)プロピオン酸tert-ブチルエステルの調製
Step 3: Preparation of (2S) -3-[(diisopropylamino) -tert-butoxyphosphinooxy] -2- (tert-butoxycarbonylamino) propionic acid tert-butyl ester
ビス(ジイソプロピルアミノ)-tert-ブチルホスフィン(643.1mg、2.112mmol)をCH2Cl2(6ml)およびトルエン(0.5ml)の混合溶媒に溶解させ、N-tert-ブトキシカルボニルセリンtert-ブチルエステル(430.1mg、1.646mmol)を加えた。水分を除去するために、得られた溶液減圧下濃縮し、アルゴン雰囲気下でCH2Cl2(6ml)を加え、THF(6ml)中の1H-テトラゾール(137.2mg、1.959mmol)を室温で加えた。数分後に白色固体が析出した。反応混合物を3時間室温で攪拌し、飽和NaHCO3水溶液(20ml)を加えて反応をクエンチし、CH2Cl2(10ml×3)で抽出した。有機層を合わせて、食塩水で洗浄し、Na2SO4で乾燥させ、濃縮した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt:Et3N=35:4:1)により精製し、表題の化合物(704.1mg、1.516mmol、92%、黄色油状物)を得た。なお、カラムクロマトグラフィーは、溶離液に3%(v/v)Et3Nを加えてシリカゲルを不活性化して行った。
Bis (diisopropylamino) -tert-butylphosphine (643.1 mg, 2.112 mmol) was dissolved in a mixed solvent of CH 2 Cl 2 (6 ml) and toluene (0.5 ml), and N-tert-butoxycarbonylserine tert- Butyl ester (430.1 mg, 1.646 mmol) was added. In order to remove moisture, the resulting solution was concentrated under reduced pressure, CH 2 Cl 2 (6 ml) was added under an argon atmosphere, and 1H-tetrazole (137.2 mg, 1.959 mmol) in THF (6 ml) was added at room temperature. Added in. A white solid precipitated after a few minutes. The reaction mixture was stirred for 3 h at room temperature, quenched with saturated aqueous NaHCO 3 (20 ml) and extracted with CH 2 Cl 2 (10 ml × 3). The organic layers were combined, washed with brine, dried over Na 2 SO 4 and concentrated. The residue was purified by column chromatography (hexane: AcOEt: Et 3 N = 35: 4: 1) to give the title compound (704.1 mg, 1.516 mmol, 92%, yellow oil). Column chromatography was performed by adding 3% (v / v) Et 3 N to the eluent to inactivate the silica gel.
1H-NMR(CD2Cl2):δ=5.500 (1/2H, d, J=8.28 Hz), 5.332 (1/2H, d, J=8.28 Hz), 4.218-4.138 (1H, m), 3.895 (1H, m), 3.747-3.650 (1H, m), 3.569 (2H, m), 1.443 (9H, d, J=2.56 Hz), 1.415 (9H, d, J=2.96 Hz), 1.326 (9H, d, J=14.2 Hz), 1.140 (12H, dt, J=2.44, 6.60 Hz)。
1 H-NMR (CD 2 Cl 2 ): δ = 5.500 (1 / 2H, d, J = 8.28 Hz), 5.332 (1 / 2H, d, J = 8.28 Hz), 4.218-4.138 (1H, m), 3.895 (1H, m), 3.747-3.650 (1H, m), 3.569 (2H, m), 1.443 (9H, d, J = 2.56 Hz), 1.415 (9H, d, J = 2.96 Hz), 1.326 (9H , d, J = 14.2 Hz), 1.140 (12H, dt, J = 2.44, 6.60 Hz).
工程4:3-オレオイルオキシ-1-プロパノールの調製
Step 4: Preparation of 3-oleoyloxy-1-propanol
1,3-プロパンジオール(623.3mg、8.192mmol)およびピリジン(0.2ml)をCH2Cl2(15ml)に溶解させ、CH2Cl2(5ml)中のオレオイルクロリド(623.4mg,、2.072mmol)を0℃で滴下して加えた。反応混合物を24時間攪拌し、5%KHSO4水溶液(15ml)により反応をクエンチした。有機層を分離し、5%KHSO4水溶液および食塩水で洗浄し、Na2SO4で乾燥させ、濃縮した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=4:1)により精製し、表題の化合物を得た(503.6mg、1.479mmol、71%、黄色油状物)。
1,3-propanediol (623.3 mg, 8.192 mmol) and pyridine (0.2 ml) were dissolved in CH 2 Cl 2 (15 ml) and oleoyl chloride (623.4 mg in CH 2 Cl 2 (5 ml). , 2.072 mmol) was added dropwise at 0 ° C. The reaction mixture was stirred for 24 hours and quenched with 5% KHSO 4 aqueous solution (15 ml). The organic layer was separated, washed with 5% aqueous KHSO 4 solution and brine, dried over Na 2 SO 4 and concentrated. The residue was purified by column chromatography (hexane / AcOEt = 4: 1) to give the title compound (503.6 mg, 1.479 mmol, 71%, yellow oil).
1H-NMR(CDCl3):δ=5.345 (2H, m), 4.239 (2H, t, J=6.12 Hz), 3.691 (2H, t, J=6.02 Hz), 2.311 (2H, t, J= 7.54 Hz), 2.016 (4H, m), 1.869 (2H, quintet, J=6.00 Hz), 1.621 (2H, m), 1.285 (20H, m), 0.878 (3H, t, J=6.82 Hz)。
13C-NMR(CDCl3):δ=174.35, 130.03, 129.74, 61.14, 59.25, 34.31, 31.91, 31.80, 29.77, 29.69, 29.53, 29.33 , 29.17, 29.13, 29.10, 27.23, 27.17, 24.98, 22.69, 14.13。
HRMS (ESI, [M+Na]+): 計算値 C21H40NaO3 +: 363.2870;実測値:363.2846。 1 H-NMR (CDCl 3 ): δ = 5.345 (2H, m), 4.239 (2H, t, J = 6.12 Hz), 3.691 (2H, t, J = 6.02 Hz), 2.311 (2H, t, J = 7.54 Hz), 2.016 (4H, m), 1.869 (2H, quintet, J = 6.00 Hz), 1.621 (2H, m), 1.285 (20H, m), 0.878 (3H, t, J = 6.82 Hz).
13 C-NMR (CDCl 3 ): δ = 174.35, 130.03, 129.74, 61.14, 59.25, 34.31, 31.91, 31.80, 29.77, 29.69, 29.53, 29.33, 29.17, 29.13, 29.10, 27.23, 27.17, 24.98, 22.69, 14.13 .
HRMS (ESI, [M + Na] + ): calculated C 21 H 40 NaO 3 + : 363.2870; found: 363.2846.
13C-NMR(CDCl3):δ=174.35, 130.03, 129.74, 61.14, 59.25, 34.31, 31.91, 31.80, 29.77, 29.69, 29.53, 29.33 , 29.17, 29.13, 29.10, 27.23, 27.17, 24.98, 22.69, 14.13。
HRMS (ESI, [M+Na]+): 計算値 C21H40NaO3 +: 363.2870;実測値:363.2846。 1 H-NMR (CDCl 3 ): δ = 5.345 (2H, m), 4.239 (2H, t, J = 6.12 Hz), 3.691 (2H, t, J = 6.02 Hz), 2.311 (2H, t, J = 7.54 Hz), 2.016 (4H, m), 1.869 (2H, quintet, J = 6.00 Hz), 1.621 (2H, m), 1.285 (20H, m), 0.878 (3H, t, J = 6.82 Hz).
13 C-NMR (CDCl 3 ): δ = 174.35, 130.03, 129.74, 61.14, 59.25, 34.31, 31.91, 31.80, 29.77, 29.69, 29.53, 29.33, 29.17, 29.13, 29.10, 27.23, 27.17, 24.98, 22.69, 14.13 .
HRMS (ESI, [M + Na] + ): calculated C 21 H 40 NaO 3 + : 363.2870; found: 363.2846.
工程5:(2S)-2-(tert-ブトキシカルボニルアミノ)-3-[(3-オレオイルオキシプロポキシ)-tert-ブトキシ-ホスホリルオキシ]プロピオン酸tertブチルエステルの調製
Step 5: Preparation of (2S) -2- (tert-butoxycarbonylamino) -3-[(3-oleoyloxypropoxy) -tert-butoxy-phosphoryloxy] propionic acid tertbutyl ester
(2S)-3-[(ジイソプロピルアミノ)-tert-ブトキシホスフィノオキシ]-2-(tert-ブトキシカルボニルアミノ)プロピオン酸tert-ブチルエステル(313.4mg、0.677mmol)をCH2Cl2(5ml)とトルエン(0.5ml)の混合溶媒に溶解させ、減圧下溶媒を留去した。残渣に3-オレオイルオキシ-1-プロパノール(278.1mg、0.817mmol)およびCH2Cl2(5ml)を加え、減圧下溶媒を留去した。アルゴン雰囲気下で残渣をCH2Cl2(5ml)に溶解させ、THF(5ml)中の1H-テトラゾール(147.7mg、2.11mmol)の溶液を室温で加えた。数分後に白色の固体が析出した。反応混合物を室温で3時間攪拌し、tert-ブチルヒドロペルオキシド(TBHP)デカン溶液(5.0-6.0M、0.272ml、1.36mmol)を室温で加え、さらに1.5時間室温で攪拌した。反応混合物を水(20ml)で希釈し、CH2Cl2(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、減圧下溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=6:1)により精製し、表題の化合物を得た(227.5mg、0.316mmol、47%、黄色油状物)。
(2S) -3-[(Diisopropylamino) -tert-butoxyphosphinooxy] -2- (tert-butoxycarbonylamino) propionic acid tert-butyl ester (313.4 mg, 0.677 mmol) was added to CH 2 Cl 2 ( 5 ml) and toluene (0.5 ml), and the solvent was distilled off under reduced pressure. To the residue were added 3-oleoyloxy-1-propanol (278.1 mg, 0.817 mmol) and CH 2 Cl 2 (5 ml), and the solvent was evaporated under reduced pressure. The residue was dissolved in CH 2 Cl 2 (5 ml) under an argon atmosphere and a solution of 1H-tetrazole (147.7 mg, 2.11 mmol) in THF (5 ml) was added at room temperature. A white solid precipitated after a few minutes. The reaction mixture was stirred at room temperature for 3 hours, tert-butyl hydroperoxide (TBHP) decane solution (5.0-6.0 M, 0.272 ml, 1.36 mmol) was added at room temperature, and further stirred for 1.5 hours at room temperature. did. The reaction mixture was diluted with water (20 ml) and extracted with CH 2 Cl 2 (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and the solvent was removed under reduced pressure. The residue was purified by column chromatography (hexane: AcOEt = 6: 1) to give the title compound (227.5 mg, 0.316 mmol, 47%, yellow oil).
1H-NMR(CDCl3):δ=5.485 (1H, m), 5.332 (2H, m), 4.348 (2H, m),4.223 (1H, m), 4.164 (2H, m), 4.061 (2H, m), 2.288 (2H, t, J=7.68 Hz), 1.995 (6H, m), 1.602 (2H, m), 1.470 (18H, s), 1.439 (9H, s), 1.278 (20H, m), 0.869 (3H, t, J=7.00 Hz)。
13C-NMR(CDCl3):δ=173.66, 168.34, 155.23, 129.98, 129.71, 83.64, 82.68, 82.65, 79.92, 67.37, 64.07, 64.02, 60.31, 54.46, 54.38, 34.17, 31.87, 29.79, 29.77, 29.74, 29.68, 29.58, 29.49, 29.29, 29.16, 29.12, 29.09, 28.29, 27.94, 27.19, 27.15, 24.89, 22.65, 14.08。
31P-NMR(CDCl3):δ=-5.584, -5.719。
HRMS (ESI, [M+Na]+):計算値:C37H70NNaO10P+: 742.4630;実測値:742.4658。 1 H-NMR (CDCl 3 ): δ = 5.485 (1H, m), 5.332 (2H, m), 4.348 (2H, m), 4.223 (1H, m), 4.164 (2H, m), 4.061 (2H, m), 2.288 (2H, t, J = 7.68 Hz), 1.995 (6H, m), 1.602 (2H, m), 1.470 (18H, s), 1.439 (9H, s), 1.278 (20H, m), 0.869 (3H, t, J = 7.00 Hz).
13 C-NMR (CDCl 3 ): δ = 173.66, 168.34, 155.23, 129.98, 129.71, 83.64, 82.68, 82.65, 79.92, 67.37, 64.07, 64.02, 60.31, 54.46, 54.38, 34.17, 31.87, 29.79, 29.77, 29.74 , 29.68, 29.58, 29.49, 29.29, 29.16, 29.12, 29.09, 28.29, 27.94, 27.19, 27.15, 24.89, 22.65, 14.08.
31 P-NMR (CDCl 3 ): δ = −5.584, −5.719.
HRMS (ESI, [M + Na] + ): Calculated: C 37 H 70 NNaO 10 P + : 742.4630; Found: 742.4658.
13C-NMR(CDCl3):δ=173.66, 168.34, 155.23, 129.98, 129.71, 83.64, 82.68, 82.65, 79.92, 67.37, 64.07, 64.02, 60.31, 54.46, 54.38, 34.17, 31.87, 29.79, 29.77, 29.74, 29.68, 29.58, 29.49, 29.29, 29.16, 29.12, 29.09, 28.29, 27.94, 27.19, 27.15, 24.89, 22.65, 14.08。
31P-NMR(CDCl3):δ=-5.584, -5.719。
HRMS (ESI, [M+Na]+):計算値:C37H70NNaO10P+: 742.4630;実測値:742.4658。 1 H-NMR (CDCl 3 ): δ = 5.485 (1H, m), 5.332 (2H, m), 4.348 (2H, m), 4.223 (1H, m), 4.164 (2H, m), 4.061 (2H, m), 2.288 (2H, t, J = 7.68 Hz), 1.995 (6H, m), 1.602 (2H, m), 1.470 (18H, s), 1.439 (9H, s), 1.278 (20H, m), 0.869 (3H, t, J = 7.00 Hz).
13 C-NMR (CDCl 3 ): δ = 173.66, 168.34, 155.23, 129.98, 129.71, 83.64, 82.68, 82.65, 79.92, 67.37, 64.07, 64.02, 60.31, 54.46, 54.38, 34.17, 31.87, 29.79, 29.77, 29.74 , 29.68, 29.58, 29.49, 29.29, 29.16, 29.12, 29.09, 28.29, 27.94, 27.19, 27.15, 24.89, 22.65, 14.08.
31 P-NMR (CDCl 3 ): δ = −5.584, −5.719.
HRMS (ESI, [M + Na] + ): Calculated: C 37 H 70 NNaO 10 P + : 742.4630; Found: 742.4658.
工程6:(2S)-2-アミノ-3-[(3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸の調製
Step 6: Preparation of (2S) -2-amino-3-[(3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
(2S)-2-(tert-ブトキシカルボニルアミノ)-3-[(3-オレオイルオキシプロポキシ)-tert-ブトキシ-ホスホリルオキシ]プロピオン酸tertブチルエステル(225.1mg、0.313mmol)を0℃でトリフルオロ酢酸(3ml)に溶解させ、溶液を室温で1.5時間攪拌した。反応混合物を濃縮し、粗製の目的物(164.2mg、褐色の油状物)を得た。粗生成物をカラムクロマトグラフィー(CHCl3/MeOH/AcOH=9:1:0→8:1:1)により精製し、表題の化合物を酢酸塩として得た(93.0mg、0.183mmol、59%、白色粉末)。さらに得られた酢酸塩をトリフルオロ酢酸に溶解させ、溶媒を留去し、トリフルオロ酢酸塩を得た(褐色の粉末)。
(2S) -2- (tert-butoxycarbonylamino) -3-[(3-oleoyloxypropoxy) -tert-butoxy-phosphoryloxy] propionic acid tertbutyl ester (225.1 mg, 0.313 mmol) was added at 0 ° C. And dissolved in trifluoroacetic acid (3 ml) and the solution was stirred at room temperature for 1.5 hours. The reaction mixture was concentrated to give a crude product (164.2 mg, brown oil). The crude product was purified by column chromatography (CHCl 3 / MeOH / AcOH = 9: 1: 0 → 8: 1: 1) to give the title compound as an acetate salt (93.0 mg, 0.183 mmol, 59 %, White powder). Further, the obtained acetate was dissolved in trifluoroacetic acid, and the solvent was distilled off to obtain a trifluoroacetate (brown powder).
1H-NMR(CDCl3):δ=5.362 (2H, m), 4,686 (2H, m), 4.550 (1H, m), 4.248 (2H, m), 4.106 (2H ,m), 2.398 (2H, t, J=7.52 Hz), 2.031 (6H, m), 1.613 (2H, m), 1.265 (20H, m), 0.874 (3H, m)。
31P-NMR(CDCl3):δ=-1.091。
HRMS (MALDI-TOF, [M+Na]+):計算値 C24H46NO8PNa+: 530.2853;実測値: 530.2810。
元素分析:計算値 C24H46NO8P・CF3COOH: C, 50.24; H, 7.62; N, 2.25;実測値: C, 50.52; H, 7.78; N, 2.44。
融点 164.3℃-165.2℃。 1 H-NMR (CDCl 3 ): δ = 5.362 (2H, m), 4,686 (2H, m), 4.550 (1H, m), 4.248 (2H, m), 4.106 (2H, m), 2.398 (2H, t, J = 7.52 Hz), 2.031 (6H, m), 1.613 (2H, m), 1.265 (20H, m), 0.874 (3H, m).
31 P-NMR (CDCl 3 ): δ = −1.091.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 24 H 46 NO 8 PNa + : 530.2853; Found: 530.2810.
Calcd C 24 H 46 NO 8 P · CF 3 COOH: C, 50.24; H, 7.62; N, 2.25; Found: C, 50.52; H, 7.78 ; N, 2.44.
Melting point: 164.3 ℃ -165.2 ℃.
31P-NMR(CDCl3):δ=-1.091。
HRMS (MALDI-TOF, [M+Na]+):計算値 C24H46NO8PNa+: 530.2853;実測値: 530.2810。
元素分析:計算値 C24H46NO8P・CF3COOH: C, 50.24; H, 7.62; N, 2.25;実測値: C, 50.52; H, 7.78; N, 2.44。
融点 164.3℃-165.2℃。 1 H-NMR (CDCl 3 ): δ = 5.362 (2H, m), 4,686 (2H, m), 4.550 (1H, m), 4.248 (2H, m), 4.106 (2H, m), 2.398 (2H, t, J = 7.52 Hz), 2.031 (6H, m), 1.613 (2H, m), 1.265 (20H, m), 0.874 (3H, m).
31 P-NMR (CDCl 3 ): δ = −1.091.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 24 H 46 NO 8 PNa + : 530.2853; Found: 530.2810.
Calcd C 24 H 46 NO 8 P · CF 3 COOH: C, 50.24; H, 7.62; N, 2.25; Found: C, 50.52; H, 7.78 ; N, 2.44.
Melting point: 164.3 ℃ -165.2 ℃.
実施例1と同様の手法により、以下の化合物を調製した。
The following compounds were prepared in the same manner as in Example 1.
[実施例2](2S)-2-アミノ-3-[(3-バセノイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 2] (2S) -2-Amino-3-[(3-basenoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3):δ= 5.338 (2H, m), 4.143-3.933 (7H, m), 2.275 (2H, m), 2.016-1.952 (4H, m), 1.634-1.588 (4H, m), 1.273 (23H, m), 0.879 (3H, t, J=6.24 Hz)。
31P-NMR(CDCl3):δ=-1.253。
HRMS (MALDI-TOF, [M+Na]+):計算値:C24H46NO8PNa+: 530.2853;実測値: 530.2809。
元素分析:計算値 C24H46NO8P・0.95CF3COOH: C, 50.51; H, 7.68; N, 2.27;実測値: C, 50.48; H, 7.61; N, 2.66。
融点 155.0℃-156.2℃。 1 H-NMR (CDCl 3 ): δ = 5.338 (2H, m), 4.143-3.933 (7H, m), 2.275 (2H, m), 2.016-1.952 (4H, m), 1.634-1.588 (4H, m ), 1.273 (23H, m), 0.879 (3H, t, J = 6.24 Hz).
31 P-NMR (CDCl 3 ): δ = -1.253.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 24 H 46 NO 8 PNa + : 530.2853; Found: 530.2809.
Calcd C 24 H 46 NO 8 P · 0.95CF 3 COOH: C, 50.51; H, 7.68; N, 2.27; Found: C, 50.48; H, 7.61 ; N, 2.66.
Melting point: 155.0 ℃ -156.2 ℃.
31P-NMR(CDCl3):δ=-1.253。
HRMS (MALDI-TOF, [M+Na]+):計算値:C24H46NO8PNa+: 530.2853;実測値: 530.2809。
元素分析:計算値 C24H46NO8P・0.95CF3COOH: C, 50.51; H, 7.68; N, 2.27;実測値: C, 50.48; H, 7.61; N, 2.66。
融点 155.0℃-156.2℃。 1 H-NMR (CDCl 3 ): δ = 5.338 (2H, m), 4.143-3.933 (7H, m), 2.275 (2H, m), 2.016-1.952 (4H, m), 1.634-1.588 (4H, m ), 1.273 (23H, m), 0.879 (3H, t, J = 6.24 Hz).
31 P-NMR (CDCl 3 ): δ = -1.253.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 24 H 46 NO 8 PNa + : 530.2853; Found: 530.2809.
Calcd C 24 H 46 NO 8 P · 0.95
Melting point: 155.0 ℃ -156.2 ℃.
[実施例3](2S)-2-アミノ-3-[(3-エライドイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 3] (2S) -2-Amino-3-[(3-Elideyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3):δ= 5.421(2H, m),4.738 (2H, m), 4.627 (1H, m), 4.296 (2H, m), 4.211 (2H, m), 2.427 (2H, t, J=7.68 Hz), 2.089(2H, m), 1.987 (4H, m), 1.655 (2H, m), 1.313 (20H, m), 0.879 (3H, t, J= 6.76 Hz)。
31P-NMR(CDCl3):δ=-1.026。
HRMS (MALDI-TOF, [M+Na]+):計算値:C24H46NNaO8P+: 530.2853;実測値:530.2823.
元素分析:計算値 C24H46NO8P・1.1CF3COOH: C, 49.71; H, 7.70; N, 2.21;実測値:C, 49.73; H, 7.72; N, 2.16。
融点 112.3℃-113.8℃。 1 H-NMR (CDCl 3 ): δ = 5.421 (2H, m), 4.738 (2H, m), 4.627 (1H, m), 4.296 (2H, m), 4.211 (2H, m), 2.427 (2H, t, J = 7.68 Hz), 2.089 (2H, m), 1.987 (4H, m), 1.655 (2H, m), 1.313 (20H, m), 0.879 (3H, t, J = 6.76 Hz).
31 P-NMR (CDCl 3 ): δ = -1.026.
HRMS (MALDI-TOF, [M + Na] + ): Calculated value: C 24 H 46 NNaO 8 P + : 530.2853; Found: 530.2823.
Calcd C 24 H 46 NO 8 P · 1.1CF 3 COOH: C, 49.71; H, 7.70; N, 2.21; Found: C, 49.73; H, 7.72 ; N, 2.16.
Melting point 112.3 ℃ -113.8 ℃.
31P-NMR(CDCl3):δ=-1.026。
HRMS (MALDI-TOF, [M+Na]+):計算値:C24H46NNaO8P+: 530.2853;実測値:530.2823.
元素分析:計算値 C24H46NO8P・1.1CF3COOH: C, 49.71; H, 7.70; N, 2.21;実測値:C, 49.73; H, 7.72; N, 2.16。
融点 112.3℃-113.8℃。 1 H-NMR (CDCl 3 ): δ = 5.421 (2H, m), 4.738 (2H, m), 4.627 (1H, m), 4.296 (2H, m), 4.211 (2H, m), 2.427 (2H, t, J = 7.68 Hz), 2.089 (2H, m), 1.987 (4H, m), 1.655 (2H, m), 1.313 (20H, m), 0.879 (3H, t, J = 6.76 Hz).
31 P-NMR (CDCl 3 ): δ = -1.026.
HRMS (MALDI-TOF, [M + Na] + ): Calculated value: C 24 H 46 NNaO 8 P + : 530.2853; Found: 530.2823.
Calcd C 24 H 46 NO 8 P · 1.1
Melting point 112.3 ℃ -113.8 ℃.
[実施例4](2S)-2-アミノ-3-[(3-パルミトレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 4] (2S) -2-Amino-3-[(3-palmitoleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3):δ= 5.375 (1H, m), 5.114 (1H, m), 4.728 (2H, m), 4.613 (1H, m), 4.286 (2H, t, J=5.98 Hz), 4.206 (2H, m), 2.418 (2H, t, J=6.88 Hz), 2.081-2.029 (4H, m), 1.681-1.634 (4H, m), 1.313 (16H, m), 0.881 (3H, t, J=6.02 Hz)。
31P-NMR(CDCl3):δ=-1.087。
HRMS (MALDI-TOF, [M+Na]+): 計算値 C22H42NO8PNa+:502.2540;実測値:502.2578.
元素分析:計算値:C22H42NO8P・1.3CF3COOH: C, 47.07; H, 6.95; N, 2.23;実測値:C, 46.74; H, 6.97; N, 2.50。
融点 151.5℃-153.0℃。 1 H-NMR (CDCl 3 ): δ = 5.375 (1H, m), 5.114 (1H, m), 4.728 (2H, m), 4.613 (1H, m), 4.286 (2H, t, J = 5.98 Hz) , 4.206 (2H, m), 2.418 (2H, t, J = 6.88 Hz), 2.081-2.029 (4H, m), 1.681-1.634 (4H, m), 1.313 (16H, m), 0.881 (3H, t , J = 6.02 Hz).
31 P-NMR (CDCl 3 ): δ = −1.087.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 22 H 42 NO 8 PNa + : 502.2540; Found: 502.2578.
Calcd: C 22 H 42 NO 8 P · 1.3CF 3 COOH: C, 47.07; H, 6.95; N, 2.23; Found: C, 46.74; H, 6.97 ; N, 2.50.
Melting point: 151.5 ° C-153.0 ° C.
31P-NMR(CDCl3):δ=-1.087。
HRMS (MALDI-TOF, [M+Na]+): 計算値 C22H42NO8PNa+:502.2540;実測値:502.2578.
元素分析:計算値:C22H42NO8P・1.3CF3COOH: C, 47.07; H, 6.95; N, 2.23;実測値:C, 46.74; H, 6.97; N, 2.50。
融点 151.5℃-153.0℃。 1 H-NMR (CDCl 3 ): δ = 5.375 (1H, m), 5.114 (1H, m), 4.728 (2H, m), 4.613 (1H, m), 4.286 (2H, t, J = 5.98 Hz) , 4.206 (2H, m), 2.418 (2H, t, J = 6.88 Hz), 2.081-2.029 (4H, m), 1.681-1.634 (4H, m), 1.313 (16H, m), 0.881 (3H, t , J = 6.02 Hz).
31 P-NMR (CDCl 3 ): δ = −1.087.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 22 H 42 NO 8 PNa + : 502.2540; Found: 502.2578.
Calcd: C 22 H 42 NO 8 P · 1.3
Melting point: 151.5 ° C-153.0 ° C.
[実施例5](2S)-2-アミノ-3-[(3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 5] (2S) -2-Amino-3-[(3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3):δ=4.728 (2H, m), 4.616 (1H, s), 4.291 (2H, m), 4.202 (2H, m), 2.421 (2H, t, J=7.54 Hz), 2.083(2H, m), 1.632 (2H, m), 1.276 (24H, m), 0.881 (3H, t, J=7.04 Hz)。
31P-NMR(CDCl3):δ=-1.088。
HRMS (MALDI-TOF, [M+Na]+): 計算値 C22H44NO8PNa+: 504.2697;実測値: 504.2683。
元素分析:計算値 C22H44NO8P・CF3COOH: C, 48.40; H, 7.83; N, 2.68;実測値: C, 48.77; H, 7.83; N, 2.68。
融点 156.5℃-157.3℃。 1 H-NMR (CDCl 3 ): δ = 4.728 (2H, m), 4.616 (1H, s), 4.291 (2H, m), 4.202 (2H, m), 2.421 (2H, t, J = 7.54 Hz) , 2.083 (2H, m), 1.632 (2H, m), 1.276 (24H, m), 0.881 (3H, t, J = 7.04 Hz).
31 P-NMR (CDCl 3 ): δ = −1.088.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 22 H 44 NO 8 PNa + : 504.2697; Found: 504.2683.
Calcd C 22 H 44 NO 8 P · CF 3 COOH: C, 48.40; H, 7.83; N, 2.68; Found: C, 48.77; H, 7.83 ; N, 2.68.
Melting point 156.5 ° C-157.3 ° C.
31P-NMR(CDCl3):δ=-1.088。
HRMS (MALDI-TOF, [M+Na]+): 計算値 C22H44NO8PNa+: 504.2697;実測値: 504.2683。
元素分析:計算値 C22H44NO8P・CF3COOH: C, 48.40; H, 7.83; N, 2.68;実測値: C, 48.77; H, 7.83; N, 2.68。
融点 156.5℃-157.3℃。 1 H-NMR (CDCl 3 ): δ = 4.728 (2H, m), 4.616 (1H, s), 4.291 (2H, m), 4.202 (2H, m), 2.421 (2H, t, J = 7.54 Hz) , 2.083 (2H, m), 1.632 (2H, m), 1.276 (24H, m), 0.881 (3H, t, J = 7.04 Hz).
31 P-NMR (CDCl 3 ): δ = −1.088.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 22 H 44 NO 8 PNa + : 504.2697; Found: 504.2683.
Calcd C 22 H 44 NO 8 P · CF 3 COOH: C, 48.40; H, 7.83; N, 2.68; Found: C, 48.77; H, 7.83 ; N, 2.68.
Melting point 156.5 ° C-157.3 ° C.
[実施例6](2S)-2-アミノ-3-[(3-ミリストイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 6] (2S) -2-Amino-3-[(3-myristoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ=4.728-4.627 (3H, m), 4.290-4.223 (4H, m), 2.421 (2H, m), 2.092 (2H, m), 1.631 (2H, m), 1.271 (20H, m), 0.878 (3H, t, J=6.80 Hz)。
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.056。
融点 152.3-153.1 ℃。
元素分析:計算値:C20H40NO8P?0.5CF3COOH: C, 49.41; H, 8.00; N, 2.74;実測値:C, 49.58; H, 7.93; N, 2.82。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 4.728-4.627 (3H, m), 4.290-4.223 (4H, m), 2.421 (2H, m), 2.092 (2H, m) , 1.631 (2H, m), 1.271 (20H, m), 0.878 (3H, t, J = 6.80 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = −1.056.
Melting point 152.3-153.1 ° C.
? Calcd: C 20 H 40 NO 8 P 0.5CF 3 COOH: C, 49.41; H, 8.00; N, 2.74; Found: C, 49.58; H, 7.93 ; N, 2.82.
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.056。
融点 152.3-153.1 ℃。
元素分析:計算値:C20H40NO8P?0.5CF3COOH: C, 49.41; H, 8.00; N, 2.74;実測値:C, 49.58; H, 7.93; N, 2.82。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 4.728-4.627 (3H, m), 4.290-4.223 (4H, m), 2.421 (2H, m), 2.092 (2H, m) , 1.631 (2H, m), 1.271 (20H, m), 0.878 (3H, t, J = 6.80 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = −1.056.
Melting point 152.3-153.1 ° C.
? Calcd: C 20 H 40 NO 8 P 0.5
[実施例7](2S)-2-アミノ-3-[(3-ラウロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 7] (2S) -2-Amino-3-[(3-lauroyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ=4.734-4.608 (3H, m), 4.286 (4H, m), 2.412 (2H, m), 2.086 (2H, m), 1.621 (2H, m), 1.264 (16H, m), 0.874 (3H, t, J=6.80 Hz)
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.299。
融点 148.1-149.0 ℃。
元素分析:計算値 C18H36NO8P?0.3CF3COOH: C, 48.60; H, 7.96; N, 3.05;実測値: C, 48.70; H, 8.24; N, 3.18。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 4.734-4.608 (3H, m), 4.286 (4H, m), 2.412 (2H, m), 2.086 (2H, m), 1.621 (2H, m), 1.264 (16H, m), 0.874 (3H, t, J = 6.80 Hz)
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.299.
Melting point 148.1-149.0 ° C.
?Calcd C 18 H 36 NO 8 P 0.3CF 3 COOH: C, 48.60; H, 7.96; N, 3.05; Found: C, 48.70; H, 8.24 ; N, 3.18.
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.299。
融点 148.1-149.0 ℃。
元素分析:計算値 C18H36NO8P?0.3CF3COOH: C, 48.60; H, 7.96; N, 3.05;実測値: C, 48.70; H, 8.24; N, 3.18。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 4.734-4.608 (3H, m), 4.286 (4H, m), 2.412 (2H, m), 2.086 (2H, m), 1.621 (2H, m), 1.264 (16H, m), 0.874 (3H, t, J = 6.80 Hz)
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.299.
Melting point 148.1-149.0 ° C.
?
[実施例8](2S)-2-アミノ-3-[(3-ヘキサデシルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 8] (2S) -2-Amino-3-[(3-hexadecyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ= 4.699 (2H, m), 4.584 (1H, m), 4.188 (2H, m), 3.771 (2H, t, J=5.94 Hz), 3.635 (2H, t, J=7.14 Hz), 2.034 (2H, m), 1.629 (2H, m), 1.264 (26H, m), 0.878 (3H, t, J=6.82 Hz)。
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.006。
元素分析:計算値 C22H46NNaO7P + 0.5CF3CO2H: C, 52.66; H, 8.93; N, 2.67;実測値:C, 52.75; H, 9.02; N, 2.76。
融点 135.1-136.4 ℃。
HRMS (MALDI-TOF, [M+Na]+):計算値:C22H46NNaO7P+:490.2904;実測値:490.2893。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 4.699 (2H, m), 4.584 (1H, m), 4.188 (2H, m), 3.771 (2H, t, J = 5.94 Hz ), 3.635 (2H, t, J = 7.14 Hz), 2.034 (2H, m), 1.629 (2H, m), 1.264 (26H, m), 0.878 (3H, t, J = 6.82 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.006.
Calcd C 22 H 46 NNaO 7 P + 0.5CF 3 CO 2 H: C, 52.66; H, 8.93; N, 2.67; Found: C, 52.75; H, 9.02 ; N, 2.76.
Melting point 135.1-136.4 ° C.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 22 H 46 NNaO 7 P + : 490.2904; Found: 490.2893.
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.006。
元素分析:計算値 C22H46NNaO7P + 0.5CF3CO2H: C, 52.66; H, 8.93; N, 2.67;実測値:C, 52.75; H, 9.02; N, 2.76。
融点 135.1-136.4 ℃。
HRMS (MALDI-TOF, [M+Na]+):計算値:C22H46NNaO7P+:490.2904;実測値:490.2893。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 4.699 (2H, m), 4.584 (1H, m), 4.188 (2H, m), 3.771 (2H, t, J = 5.94 Hz ), 3.635 (2H, t, J = 7.14 Hz), 2.034 (2H, m), 1.629 (2H, m), 1.264 (26H, m), 0.878 (3H, t, J = 6.82 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.006.
Calcd C 22 H 46 NNaO 7 P + 0.5
Melting point 135.1-136.4 ° C.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 22 H 46 NNaO 7 P + : 490.2904; Found: 490.2893.
[実施例9](2S)-2-アミノ-3-[(2-オレイルオキシエトキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 9] (2S) -2-Amino-3-[(2-oleyloxyethoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ=5.360 (2H, m), 4.630 (2H, m), 4.520 (1H, m), 4.203 (2H, m), 3.825 (2H, m), 3.665 (2H, t, J=6.44 Hz), 2.023 (4H, m), 1.621 (2H, m), 1.269 (22H, m), 0.875 (3H, t, J=6.68 Hz)。
31P-NMR(CDCl3):δ=-1.508。
融点:144.6-145.8 ℃。
元素分析:計算値:C23H46NO7P?0.75CF3COOH: C, 52.07; H, 8.34; N, 2.48;実測値:C, 51.90; H, 8.38; N, 2.56。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.360 (2H, m), 4.630 (2H, m), 4.520 (1H, m), 4.203 (2H, m), 3.825 (2H , m), 3.665 (2H, t, J = 6.44 Hz), 2.023 (4H, m), 1.621 (2H, m), 1.269 (22H, m), 0.875 (3H, t, J = 6.68 Hz).
31 P-NMR (CDCl 3 ): δ = -1.508.
Melting point: 144.6-145.8 ° C.
? Calcd: C 23 H 46 NO 7 P 0.75CF 3 COOH: C, 52.07; H, 8.34; N, 2.48; Found: C, 51.90; H, 8.38 ; N, 2.56.
31P-NMR(CDCl3):δ=-1.508。
融点:144.6-145.8 ℃。
元素分析:計算値:C23H46NO7P?0.75CF3COOH: C, 52.07; H, 8.34; N, 2.48;実測値:C, 51.90; H, 8.38; N, 2.56。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.360 (2H, m), 4.630 (2H, m), 4.520 (1H, m), 4.203 (2H, m), 3.825 (2H , m), 3.665 (2H, t, J = 6.44 Hz), 2.023 (4H, m), 1.621 (2H, m), 1.269 (22H, m), 0.875 (3H, t, J = 6.68 Hz).
31 P-NMR (CDCl 3 ): δ = -1.508.
Melting point: 144.6-145.8 ° C.
? Calcd: C 23 H 46 NO 7 P 0.75
[実施例10](2S)-2-アミノ-3-[(3-オレイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 10] (2S) -2-Amino-3-[(3-oleyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ= 5.360 (2H, m), 4.685 (2H, m), 4.569 (1H, m), 4.172 (2H, m), 3.762 (2H, m), 3.626 (2H, t, J=7.04 Hz), 2.027 (4H, m), 1.624 (4H, m), 1.262 (22H, m), 0.875 (3H, t, J=6.72 Hz)。
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.116。
HRMS (MALDI-TOF, [M+Na]+): 計算値 C24H48NNaO7P+:516.3061. Found: 516.3033。
元素分析:計算値:C24H48NO7P + 0.9CF3CO2H: C, 51.97; H, 8.27; N, 2.35;実測値:C, 52.16; H, 8.43; N, 2.49。
融点:155.1-156.1 ℃。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 5.360 (2H, m), 4.685 (2H, m), 4.569 (1H, m), 4.172 (2H, m), 3.762 (2H , m), 3.626 (2H, t, J = 7.04 Hz), 2.027 (4H, m), 1.624 (4H, m), 1.262 (22H, m), 0.875 (3H, t, J = 6.72 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.116.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 24 H 48 NNaO 7 P + : 516.3061. Found: 516.3033.
Calcd: C 24 H 48 NO 7 P + 0.9CF 3 CO 2 H: C, 51.97; H, 8.27; N, 2.35; Found: C, 52.16; H, 8.43 ; N, 2.49.
Melting point: 155.1-156.1 ° C.
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.116。
HRMS (MALDI-TOF, [M+Na]+): 計算値 C24H48NNaO7P+:516.3061. Found: 516.3033。
元素分析:計算値:C24H48NO7P + 0.9CF3CO2H: C, 51.97; H, 8.27; N, 2.35;実測値:C, 52.16; H, 8.43; N, 2.49。
融点:155.1-156.1 ℃。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 5.360 (2H, m), 4.685 (2H, m), 4.569 (1H, m), 4.172 (2H, m), 3.762 (2H , m), 3.626 (2H, t, J = 7.04 Hz), 2.027 (4H, m), 1.624 (4H, m), 1.262 (22H, m), 0.875 (3H, t, J = 6.72 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.116.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 24 H 48 NNaO 7 P + : 516.3061. Found: 516.3033.
Calcd: C 24 H 48 NO 7 P + 0.9
Melting point: 155.1-156.1 ° C.
[実施例11](2S)-2-アミノ-3-[(4-オレイルオキシブトキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 11] (2S) -2-Amino-3-[(4-oleyloxybutoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ= 5.370 (2H, m), 4.703 (2H, m), 4.584 (1H, m), 4.124 (2H, m), 3.677 (2H, m), 3.632 (2H, t, J=7.12 Hz), 2.033 (2H, m), 1.752 (4H, m), 1.625 (4H, m), 1.271 (22H, m), 0.878 (3H, t, J=6.72 Hz)。
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.140。
HRMS (MALDI-TOF, [M+Na]+):計算値 C25H50NNaO7P+:530.3217. Found: 530.3180。
元素分析:計算値 C25H50NO7P + 1.5CF3CO2H: C, 49.55; H, 7.65; N, 2.06;実測値:C, 49.43; H, 7.95; N, 2.32。
融点:158.9-159.8 ℃。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 5.370 (2H, m), 4.703 (2H, m), 4.584 (1H, m), 4.124 (2H, m), 3.677 (2H , m), 3.632 (2H, t, J = 7.12 Hz), 2.033 (2H, m), 1.752 (4H, m), 1.625 (4H, m), 1.271 (22H, m), 0.878 (3H, t, J = 6.72 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.140.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 25 H 50 NNaO 7 P + : 530.3217. Found: 530.3180.
Calcd C 25 H 50 NO 7 P + 1.5CF 3 CO 2 H: C, 49.55; H, 7.65; N, 2.06; Found: C, 49.43; H, 7.95 ; N, 2.32.
Melting point: 158.9-159.8 ° C.
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.140。
HRMS (MALDI-TOF, [M+Na]+):計算値 C25H50NNaO7P+:530.3217. Found: 530.3180。
元素分析:計算値 C25H50NO7P + 1.5CF3CO2H: C, 49.55; H, 7.65; N, 2.06;実測値:C, 49.43; H, 7.95; N, 2.32。
融点:158.9-159.8 ℃。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 5.370 (2H, m), 4.703 (2H, m), 4.584 (1H, m), 4.124 (2H, m), 3.677 (2H , m), 3.632 (2H, t, J = 7.12 Hz), 2.033 (2H, m), 1.752 (4H, m), 1.625 (4H, m), 1.271 (22H, m), 0.878 (3H, t, J = 6.72 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.140.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 25 H 50 NNaO 7 P + : 530.3217. Found: 530.3180.
Calcd C 25 H 50 NO 7 P + 1.5
Melting point: 158.9-159.8 ° C.
[実施例12](2S)-2-アミノ-3-[(5-オレイルオキシペントキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 12] (2S) -2-Amino-3-[(5-oleyloxypentoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ=5.370 (2H, m), 4.683 (2H, m), 4.562 (1H, m), 4.082 (2H, m), 3.625 (4H, m), 2.025 (4H, m), 1.665 (6H, m), 1.424 (2H, m), 1.299 (22H, m), 0.878 (3H, t, J=6.78 Hz)。
31P-NMR(CDCl3):δ=-1.121。
融点 148.2-149.4 ℃。
元素分析:計算値 C26H52NO7P?0.6CF3COOH: C, 55.36; H, 8.98; N, 2.37;実測値: C, 55.58; H, 9.16; N, 2.40。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.370 (2H, m), 4.683 (2H, m), 4.562 (1H, m), 4.082 (2H, m), 3.625 (4H , m), 2.025 (4H, m), 1.665 (6H, m), 1.424 (2H, m), 1.299 (22H, m), 0.878 (3H, t, J = 6.78 Hz).
31 P-NMR (CDCl 3 ): δ = -1.121.
Melting point 148.2-149.4 ° C.
?Calcd C 26 H 52 NO 7 P 0.6CF 3 COOH: C, 55.36; H, 8.98; N, 2.37; Found: C, 55.58; H, 9.16 ; N, 2.40.
31P-NMR(CDCl3):δ=-1.121。
融点 148.2-149.4 ℃。
元素分析:計算値 C26H52NO7P?0.6CF3COOH: C, 55.36; H, 8.98; N, 2.37;実測値: C, 55.58; H, 9.16; N, 2.40。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.370 (2H, m), 4.683 (2H, m), 4.562 (1H, m), 4.082 (2H, m), 3.625 (4H , m), 2.025 (4H, m), 1.665 (6H, m), 1.424 (2H, m), 1.299 (22H, m), 0.878 (3H, t, J = 6.78 Hz).
31 P-NMR (CDCl 3 ): δ = -1.121.
Melting point 148.2-149.4 ° C.
?
[実施例13](2S)-2-アミノ-3-[3-{8-(4-ヘプチル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 13] (2S) -2-Amino-3- [3- {8- (4-heptyl-1,2,3-triazol-1-yl) octanoyloxy} propoxy-hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3):δ= 7.878 (1H, s), 4.707 (2H, m), 4.621 (1H, m), 4.538 (2H, t, J=7.26 Hz), 4.270 (2H, m), 4.188 (2H, m), 2.874 (2H, t, J=7.86 Hz), 2.418 (2H, t, J=7.26 Hz), 2.037 (4H, m), 1.720 (2H, m), 1.628 (2H, m), 1.367-1.276 (14H, m), 0.866 (3H, t, J=6.78 Hz)。
31P-NMR(CDCl3):δ=-1.142。
HRMS (MALDI-TOF, [M+Na]+):計算値:C23H43N4NaO8P+:557.2711;実測値:557.2755。
融点 107.8-108.5℃。
元素分析:計算値 C23H43N4O8P・0.9CF3COOH: C, 46.75; H, 6.94; N, 8.79;実測値: C, 46.93; H, 7.20; N, 8.39。 1 H-NMR (CDCl 3 ): δ = 7.878 (1H, s), 4.707 (2H, m), 4.621 (1H, m), 4.538 (2H, t, J = 7.26 Hz), 4.270 (2H, m) , 4.188 (2H, m), 2.874 (2H, t, J = 7.86 Hz), 2.418 (2H, t, J = 7.26 Hz), 2.037 (4H, m), 1.720 (2H, m), 1.628 (2H, m), 1.367-1.276 (14H, m), 0.866 (3H, t, J = 6.78 Hz).
31 P-NMR (CDCl 3 ): δ = -1.142.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 23 H 43 N 4 NaO 8 P + : 557.2711; Found: 557.2755.
Melting point 107.8-108.5 ° C.
Calcd C 23 H 43 N 4 O 8 P · 0.9CF 3 COOH: C, 46.75; H, 6.94; N, 8.79; Found: C, 46.93; H, 7.20 ; N, 8.39.
31P-NMR(CDCl3):δ=-1.142。
HRMS (MALDI-TOF, [M+Na]+):計算値:C23H43N4NaO8P+:557.2711;実測値:557.2755。
融点 107.8-108.5℃。
元素分析:計算値 C23H43N4O8P・0.9CF3COOH: C, 46.75; H, 6.94; N, 8.79;実測値: C, 46.93; H, 7.20; N, 8.39。 1 H-NMR (CDCl 3 ): δ = 7.878 (1H, s), 4.707 (2H, m), 4.621 (1H, m), 4.538 (2H, t, J = 7.26 Hz), 4.270 (2H, m) , 4.188 (2H, m), 2.874 (2H, t, J = 7.86 Hz), 2.418 (2H, t, J = 7.26 Hz), 2.037 (4H, m), 1.720 (2H, m), 1.628 (2H, m), 1.367-1.276 (14H, m), 0.866 (3H, t, J = 6.78 Hz).
31 P-NMR (CDCl 3 ): δ = -1.142.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 23 H 43 N 4 NaO 8 P + : 557.2711; Found: 557.2755.
Melting point 107.8-108.5 ° C.
Calcd C 23 H 43 N 4 O 8 P · 0.9
[実施例14](2S)-2-アミノ-3-[3-{8-(4-ヘキシル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 14] (2S) -2-Amino-3- [3- {8- (4-hexyl-1,2,3-triazol-1-yl) octanoyloxy} propoxy-hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3):δ= 7.905 (1H, s), 4.743 (2H, m), 4.652 (1H, m), 4.557 (2H, t, J=7.32 Hz), 4.295 (2H, t, J=5.93 Hz), 4.214 (2H, m), 2.895 (2H, t, J=7.90 Hz), 2.440 (2H, t, J=7.50 Hz), 2.096-2.020 (4H, m), 1.737 (2H, m), 1.650 (2H, m), 1.389-1.302 (12H, m), 0.885 (3H, t, J=6.98 Hz)。
31P-NMR(CDCl3):δ=-1.054。
HRMS (MALDI-TOF, [M+Na]+):計算値 C22H41N4O8PNa+:543.2554;実測値:543.2545。
融点 113.1-114.2℃。
元素分析:計算値 C22H41N4O8P・1.25CF3COOH: C, 44.14; H, 6.45; N, 8.40;実測値: C, 43.92; H, 6.85; N, 8.67。 1 H-NMR (CDCl 3 ): δ = 7.905 (1H, s), 4.743 (2H, m), 4.652 (1H, m), 4.557 (2H, t, J = 7.32 Hz), 4.295 (2H, t, J = 5.93 Hz), 4.214 (2H, m), 2.895 (2H, t, J = 7.90 Hz), 2.440 (2H, t, J = 7.50 Hz), 2.096-2.020 (4H, m), 1.737 (2H, m), 1.650 (2H, m), 1.389-1.302 (12H, m), 0.885 (3H, t, J = 6.98 Hz).
31 P-NMR (CDCl 3 ): δ = −1.054.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 22 H 41 N 4 O 8 PNa + : 543.2554; Found: 543.2545.
Melting point 113.1-114.2 ° C.
Calcd C 22 H 41 N 4 O 8 P · 1.25CF 3 COOH: C, 44.14; H, 6.45; N, 8.40; Found: C, 43.92; H, 6.85 ; N, 8.67.
31P-NMR(CDCl3):δ=-1.054。
HRMS (MALDI-TOF, [M+Na]+):計算値 C22H41N4O8PNa+:543.2554;実測値:543.2545。
融点 113.1-114.2℃。
元素分析:計算値 C22H41N4O8P・1.25CF3COOH: C, 44.14; H, 6.45; N, 8.40;実測値: C, 43.92; H, 6.85; N, 8.67。 1 H-NMR (CDCl 3 ): δ = 7.905 (1H, s), 4.743 (2H, m), 4.652 (1H, m), 4.557 (2H, t, J = 7.32 Hz), 4.295 (2H, t, J = 5.93 Hz), 4.214 (2H, m), 2.895 (2H, t, J = 7.90 Hz), 2.440 (2H, t, J = 7.50 Hz), 2.096-2.020 (4H, m), 1.737 (2H, m), 1.650 (2H, m), 1.389-1.302 (12H, m), 0.885 (3H, t, J = 6.98 Hz).
31 P-NMR (CDCl 3 ): δ = −1.054.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 22 H 41 N 4 O 8 PNa + : 543.2554; Found: 543.2545.
Melting point 113.1-114.2 ° C.
Calcd C 22 H 41 N 4 O 8 P · 1.25
[実施例15](2S)-2-アミノ-3-[6-{4-ヘキシルオキシ-(2Z)-2-ブテン-1-イルオキシ}ヘキサノイルオキシ]プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 15] (2S) -2-Amino-3- [6- {4-hexyloxy- (2Z) -2-buten-1-yloxy} hexanoyloxy] propoxy-hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3):δ= 5.823 (2H, m), 4.714 (2H, m), 4.602 (1H, m), 4.284 (2H, t, J=5.8 Hz), 4.240 (4H, d, J=5.6 Hz), 4.186 (2H, m), 3.647 (4H, t, J=7.0 Hz), 2.430 (2H, t, J=7.6 Hz), 2.067 (2H, m), 1.698-1.605 (6H, m), 1.414-1.297 (8H, m), 0.878(3H, t, J=6.5 Hz)。
31P-NMR(CDCl3):δ=-1.144。
HRMS (MALDI-TOF, [M+Na]+):計算値 C22H42NNaO10P+: 534.2439;実測値: 534.2394。
融点 144.2-145.2℃。
元素分析:計算値:C22H42NO10P・0.7CF3COOH: C, 47.53; H, 7.28; N, 2.37;実測値:C, 47.44; H, 7.64; N, 2.66。 1 H-NMR (CDCl 3 ): δ = 5.823 (2H, m), 4.714 (2H, m), 4.602 (1H, m), 4.284 (2H, t, J = 5.8 Hz), 4.240 (4H, d, J = 5.6 Hz), 4.186 (2H, m), 3.647 (4H, t, J = 7.0 Hz), 2.430 (2H, t, J = 7.6 Hz), 2.067 (2H, m), 1.698-1.605 (6H, m), 1.414-1.297 (8H, m), 0.878 (3H, t, J = 6.5 Hz).
31 P-NMR (CDCl 3 ): δ = -1.144.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 22 H 42 NNaO 10 P + : 534.2439; Found: 534.2394.
Melting point: 144.2-145.2 ° C.
Calcd: C 22 H 42 NO 10 P · 0.7CF 3 COOH: C, 47.53; H, 7.28; N, 2.37; Found: C, 47.44; H, 7.64 ; N, 2.66.
31P-NMR(CDCl3):δ=-1.144。
HRMS (MALDI-TOF, [M+Na]+):計算値 C22H42NNaO10P+: 534.2439;実測値: 534.2394。
融点 144.2-145.2℃。
元素分析:計算値:C22H42NO10P・0.7CF3COOH: C, 47.53; H, 7.28; N, 2.37;実測値:C, 47.44; H, 7.64; N, 2.66。 1 H-NMR (CDCl 3 ): δ = 5.823 (2H, m), 4.714 (2H, m), 4.602 (1H, m), 4.284 (2H, t, J = 5.8 Hz), 4.240 (4H, d, J = 5.6 Hz), 4.186 (2H, m), 3.647 (4H, t, J = 7.0 Hz), 2.430 (2H, t, J = 7.6 Hz), 2.067 (2H, m), 1.698-1.605 (6H, m), 1.414-1.297 (8H, m), 0.878 (3H, t, J = 6.5 Hz).
31 P-NMR (CDCl 3 ): δ = -1.144.
HRMS (MALDI-TOF, [M + Na] + ): Calculated C 22 H 42 NNaO 10 P + : 534.2439; Found: 534.2394.
Melting point: 144.2-145.2 ° C.
Calcd: C 22 H 42 NO 10 P · 0.7
[実施例16](2S)-2-アミノ-3-[2-(オレイルオキシカルボニル)エトキシ-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 16] (2S) -2-Amino-3- [2- (oleyloxycarbonyl) ethoxy-hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ=5.377 (2H, m), 4.709 (2H, m), 4.609 (1H, m), 4.360 (2H, m), 4.165 (2H, m), 2.814 (2H, m), 2.033 (4H, m), 1.677 (2H, m), 1.272 (22H, m), 0.878 (3H, t, J=6.60 Hz)。
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.483。
融点:137.9-139.0 ℃。
元素分析:計算値:C24H46NO8P?0.8CF3COOH: C, 51.35; H, 7.88; N, 2.34;実測値:C, 51.42; H, 7.98; N, 2.47。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.377 (2H, m), 4.709 (2H, m), 4.609 (1H, m), 4.360 (2H, m), 4.165 (2H , m), 2.814 (2H, m), 2.033 (4H, m), 1.677 (2H, m), 1.272 (22H, m), 0.878 (3H, t, J = 6.60 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = −1.483.
Melting point: 137.9-139.0 ° C.
? Calcd: C 24 H 46 NO 8 P 0.8CF 3 COOH: C, 51.35; H, 7.88; N, 2.34; Found: C, 51.42; H, 7.98 ; N, 2.47.
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.483。
融点:137.9-139.0 ℃。
元素分析:計算値:C24H46NO8P?0.8CF3COOH: C, 51.35; H, 7.88; N, 2.34;実測値:C, 51.42; H, 7.98; N, 2.47。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.377 (2H, m), 4.709 (2H, m), 4.609 (1H, m), 4.360 (2H, m), 4.165 (2H , m), 2.814 (2H, m), 2.033 (4H, m), 1.677 (2H, m), 1.272 (22H, m), 0.878 (3H, t, J = 6.60 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = −1.483.
Melting point: 137.9-139.0 ° C.
? Calcd: C 24 H 46 NO 8 P 0.8
[実施例17](2S)-2-アミノ-3-[3-(オレイルオキシカルボニル)プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 17] (2S) -2-Amino-3- [3- (oleyloxycarbonyl) propoxy-hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ=5.368 (2H, m), 4.664 (2H, m), 4.553 (1H, m), 4.137 (4H, m), 2.528 (2H, m), 2.031 (6H, m), 1.660 (2H, m), 1.265 (22H, m), 0.877 (3H, t, J=6.78 Hz)。
31P-NMR(CDCl3):δ=-1.401。
融点 165.7-166.8 ℃。
元素分析:計算値 C25H48NO8P?0.7CF3COOH: C, 52.72; H, 8.16; N, 2.33;実測値: C, 52.60; H, 8.42; N, 2.43。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.368 (2H, m), 4.664 (2H, m), 4.553 (1H, m), 4.137 (4H, m), 2.528 (2H m), 2.031 (6H, m), 1.660 (2H, m), 1.265 (22H, m), 0.877 (3H, t, J = 6.78 Hz).
31 P-NMR (CDCl 3 ): δ = -1.401.
Melting point 165.7-166.8 ° C.
?Calcd C 25 H 48 NO 8 P 0.7CF 3 COOH: C, 52.72; H, 8.16; N, 2.33; Found: C, 52.60; H, 8.42 ; N, 2.43.
31P-NMR(CDCl3):δ=-1.401。
融点 165.7-166.8 ℃。
元素分析:計算値 C25H48NO8P?0.7CF3COOH: C, 52.72; H, 8.16; N, 2.33;実測値: C, 52.60; H, 8.42; N, 2.43。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.368 (2H, m), 4.664 (2H, m), 4.553 (1H, m), 4.137 (4H, m), 2.528 (2H m), 2.031 (6H, m), 1.660 (2H, m), 1.265 (22H, m), 0.877 (3H, t, J = 6.78 Hz).
31 P-NMR (CDCl 3 ): δ = -1.401.
Melting point 165.7-166.8 ° C.
?
[実施例18](2S)-2-アミノ-3-[4-(オレイルオキシカルボニル)ブトキシ-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 18] (2S) -2-Amino-3- [4- (oleyloxycarbonyl) butoxy-hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ=5.372 (2H, m), 4.714 (2H, m), 4.594 (1H, m), 4.143 (4H, m), 2.462 (2H, m), 2.029 (4H, m), 1.730-1.668 (6H, m), 1.268 (22H, m), 0.878 (3H, t, J=6.68 Hz)。
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.131。
融点163.4-164.3 ℃。
元素分析:計算値 C26H50NO8P?1.3CF3COOH: C, 50.23; H, 7.56; N, 2.05;実測値: C, 50.14; H, 7.64; N, 2.28。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.372 (2H, m), 4.714 (2H, m), 4.594 (1H, m), 4.143 (4H, m), 2.462 (2H , m), 2.029 (4H, m), 1.730-1.668 (6H, m), 1.268 (22H, m), 0.878 (3H, t, J = 6.68 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.131.
Melting point 163.4-164.3 ° C.
?Calcd C 26 H 50 NO 8 P 1.3CF 3 COOH: C, 50.23; H, 7.56; N, 2.05; Found: C, 50.14; H, 7.64 ; N, 2.28.
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.131。
融点163.4-164.3 ℃。
元素分析:計算値 C26H50NO8P?1.3CF3COOH: C, 50.23; H, 7.56; N, 2.05;実測値: C, 50.14; H, 7.64; N, 2.28。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.372 (2H, m), 4.714 (2H, m), 4.594 (1H, m), 4.143 (4H, m), 2.462 (2H , m), 2.029 (4H, m), 1.730-1.668 (6H, m), 1.268 (22H, m), 0.878 (3H, t, J = 6.68 Hz).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.131.
Melting point 163.4-164.3 ° C.
?
[実施例19](2S,3S)-2-アミノ-3-[(3-オレオイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸
[Example 19] (2S, 3S) -2-Amino-3-[(3-oleoyloxypropyl) -hydroxyphosphoryloxy] butyric acid
1H-NMR(CDCl3):δ= 5.384 (2H, m), 5.138 (1H, m), 4.507 (1H, s), 4.290 (2H, t, J=6.14 Hz), 4.214 (2H, m), 2.426 (2H, t, J=7.62 Hz), 2.105-1.991 (4H, m), 1.649 (6H, m), 1.278 (21H, m), 0.881 (3H, t, J=6.32 Hz)。
31P-NMR(CDCl3):δ=-1.370。
融点:158.9-160.2 ℃。
元素分析:計算値:C25H48NO8P・1.05CF3COOH: C, 50.75; H, 7.71; N, 2.18;実測値:C, 50.56; H, 8.06; N, 2.45。
HRMS (MALDI-TOF, [M+Na]+):計算値:C25H48NNaO8P+: 544.3010;実測値:544.3049。 1 H-NMR (CDCl 3 ): δ = 5.384 (2H, m), 5.138 (1H, m), 4.507 (1H, s), 4.290 (2H, t, J = 6.14 Hz), 4.214 (2H, m) , 2.426 (2H, t, J = 7.62 Hz), 2.105-1.991 (4H, m), 1.649 (6H, m), 1.278 (21H, m), 0.881 (3H, t, J = 6.32 Hz).
31 P-NMR (CDCl 3 ): δ = -1.370.
Melting point: 158.9-160.2 ° C.
Calcd: C 25 H 48 NO 8 P · 1.05CF 3 COOH: C, 50.75; H, 7.71; N, 2.18; Found: C, 50.56; H, 8.06 ; N, 2.45.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 25 H 48 NNaO 8 P + : 544.3010; Found: 544.3049.
31P-NMR(CDCl3):δ=-1.370。
融点:158.9-160.2 ℃。
元素分析:計算値:C25H48NO8P・1.05CF3COOH: C, 50.75; H, 7.71; N, 2.18;実測値:C, 50.56; H, 8.06; N, 2.45。
HRMS (MALDI-TOF, [M+Na]+):計算値:C25H48NNaO8P+: 544.3010;実測値:544.3049。 1 H-NMR (CDCl 3 ): δ = 5.384 (2H, m), 5.138 (1H, m), 4.507 (1H, s), 4.290 (2H, t, J = 6.14 Hz), 4.214 (2H, m) , 2.426 (2H, t, J = 7.62 Hz), 2.105-1.991 (4H, m), 1.649 (6H, m), 1.278 (21H, m), 0.881 (3H, t, J = 6.32 Hz).
31 P-NMR (CDCl 3 ): δ = -1.370.
Melting point: 158.9-160.2 ° C.
Calcd: C 25 H 48 NO 8 P · 1.05
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 25 H 48 NNaO 8 P + : 544.3010; Found: 544.3049.
[実施例20](2S,3S)-2-アミノ-3-[{3-((9E)-オクタデセノイルオキシ)プロピル}-ヒドロキシホスホリルオキシ]酪酸
[Example 20] (2S, 3S) -2-amino-3-[{3-((9E) -octadecenoyloxy) propyl} -hydroxyphosphoryloxy] butyric acid
1H-NMR(CDCl3):δ= 5.415 (2H, m), 5.124 (1H, m), 4.495 (1H, m), 4.285 (2H, m), 4.205 (2H, m), 2.420 (2H, t, J=7.72 Hz), 2.087 (2H, m), 1.982 (4H, m), 1.644 (4H, m), 1.281 (21H, m), 0.880 (3H, t, J=6.80 Hz)。
31P-NMR(CDCl3):δ=-1.406。
融点:171.9-173.4℃。
元素分析:計算値:C25H48NO8P・0.6CF3COOH: C, 53.33; H, 8.30; N, 2.37;実測値:C, 53.51; H, 8.12; N, 2.24。
HRMS (MALDI-TOF, [M+Na]+) :計算値:C25H48NNaO8P+: 544.3010;実測値:544.3043。 1 H-NMR (CDCl 3 ): δ = 5.415 (2H, m), 5.124 (1H, m), 4.495 (1H, m), 4.285 (2H, m), 4.205 (2H, m), 2.420 (2H, t, J = 7.72 Hz), 2.087 (2H, m), 1.982 (4H, m), 1.644 (4H, m), 1.281 (21H, m), 0.880 (3H, t, J = 6.80 Hz).
31 P-NMR (CDCl 3 ): δ = -1.406.
Melting point: 171.9-173.4 ° C.
Calcd: C 25 H 48 NO 8 P · 0.6CF 3 COOH: C, 53.33; H, 8.30; N, 2.37; Found: C, 53.51; H, 8.12 ; N, 2.24.
HRMS (MALDI-TOF, [M + Na] + ): Calculated value: C 25 H 48 NNaO 8 P + : 544.3010; Found: 544.33043.
31P-NMR(CDCl3):δ=-1.406。
融点:171.9-173.4℃。
元素分析:計算値:C25H48NO8P・0.6CF3COOH: C, 53.33; H, 8.30; N, 2.37;実測値:C, 53.51; H, 8.12; N, 2.24。
HRMS (MALDI-TOF, [M+Na]+) :計算値:C25H48NNaO8P+: 544.3010;実測値:544.3043。 1 H-NMR (CDCl 3 ): δ = 5.415 (2H, m), 5.124 (1H, m), 4.495 (1H, m), 4.285 (2H, m), 4.205 (2H, m), 2.420 (2H, t, J = 7.72 Hz), 2.087 (2H, m), 1.982 (4H, m), 1.644 (4H, m), 1.281 (21H, m), 0.880 (3H, t, J = 6.80 Hz).
31 P-NMR (CDCl 3 ): δ = -1.406.
Melting point: 171.9-173.4 ° C.
Calcd: C 25 H 48 NO 8 P · 0.6
HRMS (MALDI-TOF, [M + Na] + ): Calculated value: C 25 H 48 NNaO 8 P + : 544.3010; Found: 544.33043.
[実施例21](2S,3S)-2-アミノ-3-[(3-ステアロイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸
[Example 21] (2S, 3S) -2-Amino-3-[(3-stearoyloxypropyl) -hydroxyphosphoryloxy] butyric acid
1H-NMR(CDCl3):δ= 5.137 (1H, m), 4.506 (1H, s), 4.290 (2H, t, J=6.28 Hz), 4.216 (2H, m), 2.426 (2H, t, J=7.72 Hz), 2.093 (2H, m), 1.650 (4H, m), 1.279 (29H, m), 0.882 (3H, t, J=6.80 Hz)。
31P-NMR(CDCl3):δ=-1.350。
HRMS (MALDI-TOF, [M+Na]+):計算値:C25H50NNaO8P+: 546.3166;実測値:546.3134。
元素分析:計算値:C25H50NO8P・0.6CF3COOH: C, 53.15; H, 8.72; N, 2.37;実測値:C, 53.11; H, 8.72; N, 2.45。
融点:161.3-162.2 ℃。 1 H-NMR (CDCl 3 ): δ = 5.137 (1H, m), 4.506 (1H, s), 4.290 (2H, t, J = 6.28 Hz), 4.216 (2H, m), 2.426 (2H, t, J = 7.72 Hz), 2.093 (2H, m), 1.650 (4H, m), 1.279 (29H, m), 0.882 (3H, t, J = 6.80 Hz).
31 P-NMR (CDCl 3 ): δ = -1.350.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 25 H 50 NNaO 8 P + : 546.3166; Found: 546.3134.
Calcd: C 25 H 50 NO 8 P · 0.6CF 3 COOH: C, 53.15; H, 8.72; N, 2.37; Found: C, 53.11; H, 8.72 ; N, 2.45.
Melting point: 161.3-162.2 ° C.
31P-NMR(CDCl3):δ=-1.350。
HRMS (MALDI-TOF, [M+Na]+):計算値:C25H50NNaO8P+: 546.3166;実測値:546.3134。
元素分析:計算値:C25H50NO8P・0.6CF3COOH: C, 53.15; H, 8.72; N, 2.37;実測値:C, 53.11; H, 8.72; N, 2.45。
融点:161.3-162.2 ℃。 1 H-NMR (CDCl 3 ): δ = 5.137 (1H, m), 4.506 (1H, s), 4.290 (2H, t, J = 6.28 Hz), 4.216 (2H, m), 2.426 (2H, t, J = 7.72 Hz), 2.093 (2H, m), 1.650 (4H, m), 1.279 (29H, m), 0.882 (3H, t, J = 6.80 Hz).
31 P-NMR (CDCl 3 ): δ = -1.350.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 25 H 50 NNaO 8 P + : 546.3166; Found: 546.3134.
Calcd: C 25 H 50 NO 8 P · 0.6
Melting point: 161.3-162.2 ° C.
[実施例22](2S,3S)-2-アミノ-3-[(3-パルミトレオイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸
[Example 22] (2S, 3S) -2-Amino-3-[(3-palmitoleoyloxypropyl) -hydroxyphosphoryloxy] butyric acid
1H-NMR(CDCl3):δ= 5.385 (2H, m), 5.133 (1H, m), 4.501 (1H, m), 4.288 (2H, m), 4.212 (2H, m), 2.422 (2H, t, J=7.66 Hz), 2.091-1.986 (4H, m), 1.657 (6H, m), 1.319 (17H, m), 0.883 (3H, t, J=6.92 Hz)。
31P-NMR(CDCl3):δ=-1.372。
融点:203.7-205.1℃。
HRMS (MALDI-TOF, [M+Na]+):計算値:C23H44NNaO8P+: 516.2697;実測値:516.2737。 1 H-NMR (CDCl 3 ): δ = 5.385 (2H, m), 5.133 (1H, m), 4.501 (1H, m), 4.288 (2H, m), 4.212 (2H, m), 2.422 (2H, t, J = 7.66 Hz), 2.091-1.986 (4H, m), 1.657 (6H, m), 1.319 (17H, m), 0.883 (3H, t, J = 6.92 Hz).
31 P-NMR (CDCl 3 ): δ = -1.372.
Melting point: 203.7-205.1 ° C.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 23 H 44 NNaO 8 P + : 516.2697; Found: 516.2737.
31P-NMR(CDCl3):δ=-1.372。
融点:203.7-205.1℃。
HRMS (MALDI-TOF, [M+Na]+):計算値:C23H44NNaO8P+: 516.2697;実測値:516.2737。 1 H-NMR (CDCl 3 ): δ = 5.385 (2H, m), 5.133 (1H, m), 4.501 (1H, m), 4.288 (2H, m), 4.212 (2H, m), 2.422 (2H, t, J = 7.66 Hz), 2.091-1.986 (4H, m), 1.657 (6H, m), 1.319 (17H, m), 0.883 (3H, t, J = 6.92 Hz).
31 P-NMR (CDCl 3 ): δ = -1.372.
Melting point: 203.7-205.1 ° C.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 23 H 44 NNaO 8 P + : 516.2697; Found: 516.2737.
[実施例23](2S,3S)-2-アミノ-3-[(3-パルミトイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸
[Example 23] (2S, 3S) -2-Amino-3-[(3-palmitoyloxypropyl) -hydroxyphosphoryloxy] butyric acid
1H-NMR(CDCl3):δ= 5.146 (1H, m), 4.518 (1H, m), 4.297 (2H, m), 4.225 (2H, m), 2.433 (2H, t, J=7.54 Hz), 2.099 (2H, m), 1.654 (4H, m), 1.285 (25H, m), 0.883 (3H, t, J=6.17 Hz)。
31P-NMR(CDCl3):δ=-1.363。
融点:153.4-154.1℃。
元素分析:計算値:C25H46NO8P・0.65CF3COOH: C, 51.23; H, 8.25; N, 2.46;実測値:C, 51.17; H, 8.52; N, 2.81。
HRMS (MALDI-TOF, [M+Na]+):計算値:C23H46NNaO8P+: 518.2853;実測値:518.2817。 1 H-NMR (CDCl 3 ): δ = 5.146 (1H, m), 4.518 (1H, m), 4.297 (2H, m), 4.225 (2H, m), 2.433 (2H, t, J = 7.54 Hz) , 2.099 (2H, m), 1.654 (4H, m), 1.285 (25H, m), 0.883 (3H, t, J = 6.17 Hz).
31 P-NMR (CDCl 3 ): δ = -1.363.
Melting point: 153.4-154.1 ° C.
Calcd: C 25 H 46 NO 8 P · 0.65CF 3 COOH: C, 51.23; H, 8.25; N, 2.46; Found: C, 51.17; H, 8.52 ; N, 2.81.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 23 H 46 NNaO 8 P + : 518.2853; Found: 518.2817.
31P-NMR(CDCl3):δ=-1.363。
融点:153.4-154.1℃。
元素分析:計算値:C25H46NO8P・0.65CF3COOH: C, 51.23; H, 8.25; N, 2.46;実測値:C, 51.17; H, 8.52; N, 2.81。
HRMS (MALDI-TOF, [M+Na]+):計算値:C23H46NNaO8P+: 518.2853;実測値:518.2817。 1 H-NMR (CDCl 3 ): δ = 5.146 (1H, m), 4.518 (1H, m), 4.297 (2H, m), 4.225 (2H, m), 2.433 (2H, t, J = 7.54 Hz) , 2.099 (2H, m), 1.654 (4H, m), 1.285 (25H, m), 0.883 (3H, t, J = 6.17 Hz).
31 P-NMR (CDCl 3 ): δ = -1.363.
Melting point: 153.4-154.1 ° C.
Calcd: C 25 H 46 NO 8 P · 0.65
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 23 H 46 NNaO 8 P + : 518.2853; Found: 518.2817.
なお、実施例9の製造において、以下の化合物を合成中間体として使用した。
In the production of Example 9, the following compounds were used as synthetic intermediates.
[製造例1]2-オレイルオキシ-1-エタノール
[Production Example 1] 2-Oleyloxy-1-ethanol
工程1:オレイルアルコールの調製
Step 1: Preparation of oleyl alcohol
MeOH(5ml)をオレオイルクロリド(303.1mg、1.007mmol)に0℃で加え、溶液を室温で20分間攪拌した。メタノールを留去し、オレイン酸メチルエステルを得た。
MeOH (5 ml) was added to oleoyl chloride (303.1 mg, 1.007 mmol) at 0 ° C. and the solution was stirred at room temperature for 20 minutes. Methanol was distilled off to obtain oleic acid methyl ester.
LiAlH4(58.1mg、1.531mmol)をTHF(5ml)に懸濁させ、THF(5ml)中のオレイン酸メチルエステル溶液を0℃で加え、室温で3時間攪拌した。反応混合物にNaSO4・10H2Oを加え、セライト(登録商標)で濾過し、濾液を濃縮した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=5:1)により精製し、表題の化合物を得た(260.7mg、0.971mmol、96%、無色油状物)。
LiAlH 4 (58.1 mg, 1.531 mmol) was suspended in THF (5 ml), a solution of oleic acid methyl ester in THF (5 ml) was added at 0 ° C., and the mixture was stirred at room temperature for 3 hours. NaSO 4 · 10H 2 O was added to the reaction mixture, the mixture was filtered through Celite (registered trademark), and the filtrate was concentrated. The residue was purified by column chromatography (hexane: AcOEt = 5: 1) to give the title compound (260.7 mg, 0.971 mmol, 96%, colorless oil).
1H-NMR(CDCl3):δ=5.347 (2H, m), 3.640 (2H, dt, J=6.52, 5.08 Hz), 2.020 (4H, m), 1.565 (2H, m), 1.303-1.268 (22H, m), 1.257 (1H, t, J=5.08 Hz), 0.881 (3H, t, J=6.86 Hz)。
1 H-NMR (CDCl 3 ): δ = 5.347 (2H, m), 3.640 (2H, dt, J = 6.52, 5.08 Hz), 2.020 (4H, m), 1.565 (2H, m), 1.303-1.268 ( 22H, m), 1.257 (1H, t, J = 5.08 Hz), 0.881 (3H, t, J = 6.86 Hz).
工程2:オレイルブロミドの調製
Step 2: Preparation of oleyl bromide
オレイルアルコール(240.5mg、0.896mmol)およびCBr4(345.1mg、1.041mmol)をCH2Cl2(5ml)に溶解させ、CH2Cl2(5ml)中のPPh3(345.3mg、1.316mmol)の溶液を0℃で加え、室温で30分間攪拌した。反応混合物を濃縮し、残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=10:1)により精製し、目的の化合物を得た(296.0mg、0.893mmol、定量的、無色の油状物)。
Oleyl alcohol (240.5mg, 0.896mmol) and CBr 4 (345.1mg, 1.041mmol) was dissolved in CH 2 Cl 2 (5ml), PPh 3 in CH 2 Cl 2 (5ml) ( 345.3mg , 1.316 mmol) at 0 ° C. and stirred at room temperature for 30 minutes. The reaction mixture was concentrated, and the residue was purified by column chromatography (hexane: AcOEt = 10: 1) to obtain the target compound (296.0 mg, 0.893 mmol, quantitative, colorless oil).
1H-NMR(CDCl3):δ=5.349 (2H, m), 3.406 (2H, t, J=6.88 Hz), 2.015 (4H, m), 1.854 (2H, m), 1.424 (2H, m), 1.303 (20H, m), 0.882 (3H, t, J=6.82 Hz)。
1 H-NMR (CDCl 3 ): δ = 5.349 (2H, m), 3.406 (2H, t, J = 6.88 Hz), 2.015 (4H, m), 1.854 (2H, m), 1.424 (2H, m) , 1.303 (20H, m), 0.882 (3H, t, J = 6.82 Hz).
工程3:2-オレイルオキシ-1-エタノールの調製
Step 3: Preparation of 2-oleyloxy-1-ethanol
NaH(549.2mg、13.73mmol)をDMF(5ml)中に懸濁させ、0℃で撹拌し、DMF(5ml)中のエチレングリコール(1.1255mg、18.13mmol)の溶液を滴下して加え、0℃で30分間攪拌した。THF(10ml)中のオレイルブロミド(1.5002g、4.530mmol)の溶液を滴下して加え、反応混合物を14時間攪拌した。反応混合物をH2O(10ml)で希釈し、Et2O(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=6:1)により精製し、表題の化合物を得た(954.4mg、3.054mmol、67%、無色の油状物)。
NaH (549.2 mg, 13.73 mmol) was suspended in DMF (5 ml), stirred at 0 ° C. and a solution of ethylene glycol (1.1255 mg, 18.13 mmol) in DMF (5 ml) was added dropwise. In addition, the mixture was stirred at 0 ° C. for 30 minutes. A solution of oleyl bromide (1.5002 g, 4.530 mmol) in THF (10 ml) was added dropwise and the reaction mixture was stirred for 14 hours. The reaction mixture was diluted with H 2 O (10 ml) and extracted with Et 2 O (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 6: 1) to give the title compound (954.4 mg, 3.054 mmol, 67%, colorless oil).
1H NMR (CDCl3) :δ=5.345 (2H, m), 3.726 (2H, m), 3.529 (2H, t, J=4.64 Hz), 3.467 (2H, t, J=6.70 Hz), 1.999 (5H, m), 1.585 (2H, m), 1.299-1.266 (22H, m), 0.879 (3H, t, J=6.84 Hz)。
13C NMR (CDCl3) :δ=129.95, 129.81, 71.66, 71.41, 61.88, 31.90, 29.76, 29.74, 29.68, 29.66, 29.51, 29.48, 29.45, 29.34, 29.31, 29.24, 27.20, 26.11, 22.67, 14.10。
HRMS (ESI, [M+Na]+):計算値 C20H40NaO2 +: 335.2921;実測値: 335.2910。 1 H NMR (CDCl 3 ): δ = 5.345 (2H, m), 3.726 (2H, m), 3.529 (2H, t, J = 4.64 Hz), 3.467 (2H, t, J = 6.70 Hz), 1.999 ( 5H, m), 1.585 (2H, m), 1.299-1.266 (22H, m), 0.879 (3H, t, J = 6.84 Hz).
13 C NMR (CDCl 3 ): δ = 129.95, 129.81, 71.66, 71.41, 61.88, 31.90, 29.76, 29.74, 29.68, 29.66, 29.51, 29.48, 29.45, 29.34, 29.31, 29.24, 27.20, 26.11, 22.67, 14.10.
HRMS (ESI, [M + Na] + ): Calculated C 20 H 40 NaO 2 + : 335.2921; Found: 335.2910.
13C NMR (CDCl3) :δ=129.95, 129.81, 71.66, 71.41, 61.88, 31.90, 29.76, 29.74, 29.68, 29.66, 29.51, 29.48, 29.45, 29.34, 29.31, 29.24, 27.20, 26.11, 22.67, 14.10。
HRMS (ESI, [M+Na]+):計算値 C20H40NaO2 +: 335.2921;実測値: 335.2910。 1 H NMR (CDCl 3 ): δ = 5.345 (2H, m), 3.726 (2H, m), 3.529 (2H, t, J = 4.64 Hz), 3.467 (2H, t, J = 6.70 Hz), 1.999 ( 5H, m), 1.585 (2H, m), 1.299-1.266 (22H, m), 0.879 (3H, t, J = 6.84 Hz).
13 C NMR (CDCl 3 ): δ = 129.95, 129.81, 71.66, 71.41, 61.88, 31.90, 29.76, 29.74, 29.68, 29.66, 29.51, 29.48, 29.45, 29.34, 29.31, 29.24, 27.20, 26.11, 22.67, 14.10.
HRMS (ESI, [M + Na] + ): Calculated C 20 H 40 NaO 2 + : 335.2921; Found: 335.2910.
同様の手法により、以下の製造例2~4の化合物を調製し、それぞれ実施例10~12の化合物の製造に使用した。
In the same manner, the following compounds of Production Examples 2 to 4 were prepared and used for production of the compounds of Examples 10 to 12, respectively.
[製造例2]3-オレイルオキシ-1-プロパノール
[Production Example 2] 3-Oleyloxy-1-propanol
1H NMR (CDCl3) :δ=5.345 (2H, m), 3.778 (2H, t, J=5.55 Hz), 3.612 (2H, t, J=5.70 Hz), 3.424 (2H, t, J=6.64 Hz), 2.472 (1H, brs), 2.010 (4H, m), 1.830 (2H, m), 1.564 (2H, m), 1.279 (22H, m), 0.880 (3H, t, J=6.86 Hz)。
13C NMR (CDCl3) :δ=129.94, 129.84, 71.50, 70.48, 62.51, 31.94, 31.90, 29.77, 29.75, 29.69, 29.52, 29.46, 29.43, 29.31, 29.25, 27.21, 27.20, 26.14, 22.67, 14.10。
HRMS (ESI, [M+Na]+):計算値:C21H42NaO2 +: 349.3077. Found: 349.3070。 1 H NMR (CDCl 3 ): δ = 5.345 (2H, m), 3.778 (2H, t, J = 5.55 Hz), 3.612 (2H, t, J = 5.70 Hz), 3.424 (2H, t, J = 6.64 Hz), 2.472 (1H, brs), 2.010 (4H, m), 1.830 (2H, m), 1.564 (2H, m), 1.279 (22H, m), 0.880 (3H, t, J = 6.86 Hz).
13 C NMR (CDCl 3 ): δ = 129.94, 129.84, 71.50, 70.48, 62.51, 31.94, 31.90, 29.77, 29.75, 29.69, 29.52, 29.46, 29.43, 29.31, 29.25, 27.21, 27.20, 26.14, 22.67, 14.10.
HRMS (ESI, [M + Na] + ): Calculated: C 21 H 42 NaO 2 + : 349.3077. Found: 349.3070.
13C NMR (CDCl3) :δ=129.94, 129.84, 71.50, 70.48, 62.51, 31.94, 31.90, 29.77, 29.75, 29.69, 29.52, 29.46, 29.43, 29.31, 29.25, 27.21, 27.20, 26.14, 22.67, 14.10。
HRMS (ESI, [M+Na]+):計算値:C21H42NaO2 +: 349.3077. Found: 349.3070。 1 H NMR (CDCl 3 ): δ = 5.345 (2H, m), 3.778 (2H, t, J = 5.55 Hz), 3.612 (2H, t, J = 5.70 Hz), 3.424 (2H, t, J = 6.64 Hz), 2.472 (1H, brs), 2.010 (4H, m), 1.830 (2H, m), 1.564 (2H, m), 1.279 (22H, m), 0.880 (3H, t, J = 6.86 Hz).
13 C NMR (CDCl 3 ): δ = 129.94, 129.84, 71.50, 70.48, 62.51, 31.94, 31.90, 29.77, 29.75, 29.69, 29.52, 29.46, 29.43, 29.31, 29.25, 27.21, 27.20, 26.14, 22.67, 14.10.
HRMS (ESI, [M + Na] + ): Calculated: C 21 H 42 NaO 2 + : 349.3077. Found: 349.3070.
[製造例3]4-オレイルオキシ-1-ブタノール
[Production Example 3] 4-Oleyloxy-1-butanol
1H NMR (CDCl3) :δ=5.342 (2H, m), 3.639 (2H, t, J=5.82 Hz), 3.451 (2H, t, J=5.80 Hz), 3.423 (2H, t, J=6.72 Hz), 2.524 (1H, brs), 2.007 (4H, m), 1.668 (4H, m), 1.570 (2H, m), 1.277 (22H, m), 0.878 (3H, t, J=6.86 Hz).
13C NMR (CDCl3) :δ=129.92, 129.83, 71.20, 70.88, 62.78, 31.89, 30.49, 29.76, 29.74, 29.64, 29.62, 29.51, 29.45, 29.43, 29.34, 29.31, 29.25, 27.19, 27.05, 26.13, 22.67, 14.09.
HRMS (ESI, [M+Na]+):計算値:C22H44NaO2 +: 363.3234. Found: 363.3216。 1 H NMR (CDCl 3 ): δ = 5.342 (2H, m), 3.639 (2H, t, J = 5.82 Hz), 3.451 (2H, t, J = 5.80 Hz), 3.423 (2H, t, J = 6.72 Hz), 2.524 (1H, brs), 2.007 (4H, m), 1.668 (4H, m), 1.570 (2H, m), 1.277 (22H, m), 0.878 (3H, t, J = 6.86 Hz).
13 C NMR (CDCl 3 ): δ = 129.92, 129.83, 71.20, 70.88, 62.78, 31.89, 30.49, 29.76, 29.74, 29.64, 29.62, 29.51, 29.45, 29.43, 29.34, 29.31, 29.25, 27.19, 27.05, 26.13, 22.67, 14.09.
HRMS (ESI, [M + Na] + ): Calculated: C 22 H 44 NaO 2 + : 363.3234. Found: 363.3216.
13C NMR (CDCl3) :δ=129.92, 129.83, 71.20, 70.88, 62.78, 31.89, 30.49, 29.76, 29.74, 29.64, 29.62, 29.51, 29.45, 29.43, 29.34, 29.31, 29.25, 27.19, 27.05, 26.13, 22.67, 14.09.
HRMS (ESI, [M+Na]+):計算値:C22H44NaO2 +: 363.3234. Found: 363.3216。 1 H NMR (CDCl 3 ): δ = 5.342 (2H, m), 3.639 (2H, t, J = 5.82 Hz), 3.451 (2H, t, J = 5.80 Hz), 3.423 (2H, t, J = 6.72 Hz), 2.524 (1H, brs), 2.007 (4H, m), 1.668 (4H, m), 1.570 (2H, m), 1.277 (22H, m), 0.878 (3H, t, J = 6.86 Hz).
13 C NMR (CDCl 3 ): δ = 129.92, 129.83, 71.20, 70.88, 62.78, 31.89, 30.49, 29.76, 29.74, 29.64, 29.62, 29.51, 29.45, 29.43, 29.34, 29.31, 29.25, 27.19, 27.05, 26.13, 22.67, 14.09.
HRMS (ESI, [M + Na] + ): Calculated: C 22 H 44 NaO 2 + : 363.3234. Found: 363.3216.
[製造例4]5-オレイルオキシ-1-ペンタノール
[Production Example 4] 5-Oleyloxy-1-pentanol
1H NMR (CDCl3) :δ=5.342 (2H, m), 3.646 (2H, t, J=6.50 Hz), 3.407 (2H, t, J=6.56 Hz), 3.388 (2H, t, J=6.76 Hz), 2.014 (4H, m), 1.576 (6H, m), 1.424 (3H, m), 1.291 (22H, m), 0.877 (3H, t, J=6.86 Hz).
13C NMR (CDCl3) :δ=129.92, 129.84, 71.05, 70.74, 62.86, 32.52, 31.89, 29.74, 29.51, 29.49, 29.46, 29.44, 29.31, 29.25, 27.19, 26.18, 22.67, 22.45, 14.09。
HRMS (ESI, [M+Na]+):計算値:C23H46NaO2 +: 377.3390;実測値: 377.3366。 1 H NMR (CDCl 3 ): δ = 5.342 (2H, m), 3.646 (2H, t, J = 6.50 Hz), 3.407 (2H, t, J = 6.56 Hz), 3.388 (2H, t, J = 6.76 Hz), 2.014 (4H, m), 1.576 (6H, m), 1.424 (3H, m), 1.291 (22H, m), 0.877 (3H, t, J = 6.86 Hz).
13 C NMR (CDCl 3 ): δ = 129.92, 129.84, 71.05, 70.74, 62.86, 32.52, 31.89, 29.74, 29.51, 29.49, 29.46, 29.44, 29.31, 29.25, 27.19, 26.18, 22.67, 22.45, 14.09.
HRMS (ESI, [M + Na] + ): Calculated: C 23 H 46 NaO 2 + : 377.3390; Found: 377.3366.
13C NMR (CDCl3) :δ=129.92, 129.84, 71.05, 70.74, 62.86, 32.52, 31.89, 29.74, 29.51, 29.49, 29.46, 29.44, 29.31, 29.25, 27.19, 26.18, 22.67, 22.45, 14.09。
HRMS (ESI, [M+Na]+):計算値:C23H46NaO2 +: 377.3390;実測値: 377.3366。 1 H NMR (CDCl 3 ): δ = 5.342 (2H, m), 3.646 (2H, t, J = 6.50 Hz), 3.407 (2H, t, J = 6.56 Hz), 3.388 (2H, t, J = 6.76 Hz), 2.014 (4H, m), 1.576 (6H, m), 1.424 (3H, m), 1.291 (22H, m), 0.877 (3H, t, J = 6.86 Hz).
13 C NMR (CDCl 3 ): δ = 129.92, 129.84, 71.05, 70.74, 62.86, 32.52, 31.89, 29.74, 29.51, 29.49, 29.46, 29.44, 29.31, 29.25, 27.19, 26.18, 22.67, 22.45, 14.09.
HRMS (ESI, [M + Na] + ): Calculated: C 23 H 46 NaO 2 + : 377.3390; Found: 377.3366.
実施例13の製造において、以下の化合物を合成中間体として使用した。
In the production of Example 13, the following compounds were used as synthetic intermediates.
[製造例5]3-{8-(4-ヘプチル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}-1-プロパノール
[Production Example 5] 3- {8- (4-Heptyl-1,2,3-triazol-1-yl) octanoyloxy} -1-propanol
工程1:8-アジドオクタン酸の調製
N3-(CH2)7COOH
DMF(80ml)中の8-ブロモ-n-オクタン酸(4.509g、20.21mmol)の溶液にNaN3(5.76g、88.60mmol)を加え、室温で21時間攪拌した。反応混合物にKHSO4水溶液(1N、50ml)を加え、ヘキサン(100ml×3)で抽出した。合わせた有機層をで乾燥させ、溶媒を留去し、表題の化合物を得た(2.957g、15.96mmol、79%、無色油状物)。 Step 1: Preparation of 8-azidooctanoic acid N 3 — (CH 2 ) 7 COOH
To a solution of 8-bromo-n-octanoic acid (4.509 g, 20.21 mmol) in DMF (80 ml) was added NaN 3 (5.76 g, 88.60 mmol) and stirred at room temperature for 21 hours. To the reaction mixture was added KHSO 4 aqueous solution (1N, 50 ml), and the mixture was extracted with hexane (100 ml × 3). The combined organic layers were dried over and the solvent was evaporated to give the title compound (2.957 g, 15.96 mmol, 79%, colorless oil).
N3-(CH2)7COOH
DMF(80ml)中の8-ブロモ-n-オクタン酸(4.509g、20.21mmol)の溶液にNaN3(5.76g、88.60mmol)を加え、室温で21時間攪拌した。反応混合物にKHSO4水溶液(1N、50ml)を加え、ヘキサン(100ml×3)で抽出した。合わせた有機層をで乾燥させ、溶媒を留去し、表題の化合物を得た(2.957g、15.96mmol、79%、無色油状物)。 Step 1: Preparation of 8-azidooctanoic acid N 3 — (CH 2 ) 7 COOH
To a solution of 8-bromo-n-octanoic acid (4.509 g, 20.21 mmol) in DMF (80 ml) was added NaN 3 (5.76 g, 88.60 mmol) and stirred at room temperature for 21 hours. To the reaction mixture was added KHSO 4 aqueous solution (1N, 50 ml), and the mixture was extracted with hexane (100 ml × 3). The combined organic layers were dried over and the solvent was evaporated to give the title compound (2.957 g, 15.96 mmol, 79%, colorless oil).
1H NMR (CDCl3) :δ=3.254 (2H, t, J=6.80 Hz), 2.348 (2H, t, J=7.60 Hz), 1.640-1.595 (4H, m), 1.352 (6H, m)。
1 H NMR (CDCl 3 ): δ = 3.254 (2H, t, J = 6.80 Hz), 2.348 (2H, t, J = 7.60 Hz), 1.640-1.595 (4H, m), 1.352 (6H, m).
工程2:8-(4-ヘプチル-1,2,3-トリアゾル-1-イル)オクタン酸の調製
Step 2: Preparation of 8- (4-heptyl-1,2,3-triazol-1-yl) octanoic acid
8-アジドオクタン酸(1.007g、5.436mmol)および1-ノニン(678.2mg、5.460mmol)をtert-ブチルアルコールおよび水の1:1混合液(21.6ml)中に懸濁させた。アスコルビン酸ナトリウム(0.55mmol、新たに調製した1M水溶液で550μl)、続いて水(200μl)中の硫酸銅(II)5水和物(14.9mg、0.0597mmol)の溶液を加え、混合物を23時間激しく攪拌した。反応混合物を水(60ml)で希釈し、氷冷し、発生した白色の析出物を回収した。冷水で析出物を洗浄した(10ml×2)。得られた固体を減圧下乾燥し、表題の化合物を得た(1.316g、4.253mmol、78%、白色固体)。
8-Azidooctanoic acid (1.007 g, 5.436 mmol) and 1-nonine (678.2 mg, 5.460 mmol) were suspended in a 1: 1 mixture of tert-butyl alcohol and water (21.6 ml). It was. Sodium ascorbate (0.55 mmol, 550 μl in a freshly prepared 1M aqueous solution) was added followed by a solution of copper (II) sulfate pentahydrate (14.9 mg, 0.0597 mmol) in water (200 μl) and the mixture Was stirred vigorously for 23 hours. The reaction mixture was diluted with water (60 ml) and ice-cooled, and the white precipitate generated was recovered. The precipitate was washed with cold water (10 ml × 2). The resulting solid was dried under reduced pressure to give the title compound (1.316 g, 4.253 mmol, 78%, white solid).
1H NMR (CDCl3) :δ=7.260 (1H, s), 4.304 (2H, t, J=6.80 Hz), 2.706 (2H, m), 2.401 (2H, m), 1.885 (2H, m), 1.649 (4H, m), 1.335-1.271 (14H, m), 0.874 (3H, t, J=6.80 Hz)。
HRMS (ESI, [M-H]-):計算値 C17H30N3O2 -: 308.2344;実測値:308.2344。
融点: 63.2-64.0℃。
工程3:3-{8-(4-ヘプチル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}-1-プロパノール 1 H NMR (CDCl 3 ): δ = 7.260 (1H, s), 4.304 (2H, t, J = 6.80 Hz), 2.706 (2H, m), 2.401 (2H, m), 1.885 (2H, m), 1.649 (4H, m), 1.335-1.271 (14H, m), 0.874 (3H, t, J = 6.80 Hz).
HRMS (ESI, [MH] − ): Calculated C 17 H 30 N 3 O 2 − : 308.2344; Found: 308.2344.
Melting point: 63.2-64.0 ° C.
Step 3: 3- {8- (4-Heptyl-1,2,3-triazol-1-yl) octanoyloxy} -1-propanol
HRMS (ESI, [M-H]-):計算値 C17H30N3O2 -: 308.2344;実測値:308.2344。
融点: 63.2-64.0℃。
工程3:3-{8-(4-ヘプチル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}-1-プロパノール 1 H NMR (CDCl 3 ): δ = 7.260 (1H, s), 4.304 (2H, t, J = 6.80 Hz), 2.706 (2H, m), 2.401 (2H, m), 1.885 (2H, m), 1.649 (4H, m), 1.335-1.271 (14H, m), 0.874 (3H, t, J = 6.80 Hz).
HRMS (ESI, [MH] − ): Calculated C 17 H 30 N 3 O 2 − : 308.2344; Found: 308.2344.
Melting point: 63.2-64.0 ° C.
Step 3: 3- {8- (4-Heptyl-1,2,3-triazol-1-yl) octanoyloxy} -1-propanol
1,3-プロパンジオール(300.2mg、3.945mmol)、8-(4-ヘプチル-1,2,3-トリアゾル-1-イル)オクタン酸(301.7g、0.975mmol)およびDMAP(11.4mg、0.095mmol)をCH2Cl2(10ml)に溶解させ、0℃でEDCI(194.8mg、1.016mmol)を加え、室温で14時間攪拌した。5%KHSO4水溶液(10ml)により反応をクエンチし、有機層を5%KHSO4水溶液(10ml×3)および食塩水で洗浄し、Na2SO4で乾燥させた。溶媒を留去し、残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=2:3)により精製し、表題の化合物を得た(266.2mg、0.724mmol、74%、白色固体)。
1,3-propanediol (300.2 mg, 3.945 mmol), 8- (4-heptyl-1,2,3-triazol-1-yl) octanoic acid (301.7 g, 0.975 mmol) and DMAP (11 .4 mg, 0.095 mmol) was dissolved in CH 2 Cl 2 (10 ml), EDCI (194.8 mg, 1.016 mmol) was added at 0 ° C., and the mixture was stirred at room temperature for 14 hours. The reaction was quenched with 5% KHSO 4 aqueous solution (10 ml), and the organic layer was washed with 5% KHSO 4 aqueous solution (10 ml × 3) and brine and dried over Na 2 SO 4 . The solvent was distilled off and the residue was purified by column chromatography (hexane / AcOEt = 2: 3) to give the title compound (266.2 mg, 0.724 mmol, 74%, white solid).
1H NMR (CDCl3) :δ=7.234 (1H, s), 4.298 (2H, t, J=7.16 Hz), 4.231 (2H, t, J=6.12 Hz), 3.693 (2H, t, J=6.08 Hz), 2.698 (2H, t, J=7.74 Hz), 2.311 (2H, t, J=7.44 Hz), 2.138 (1H, brs), 1.872 (4H, m), 1.621 (4H, m), 1.327-1.238 (14H, m), 0.873 (3H, t, J=6.94 Hz)。
13C NMR (CDCl3) :δ= 174.16, 148.50, 120.50, 61.37, 59.08, 50.12, 34.24, 31.82 (×2), 30.26, 29.54, 29.28, 29.10, 28.84, 28.59, 26.26, 25.72, 24.80, 22.70, 14.16。
HRMS (ESI, [M+Na]+):計算値 C20H37N3NaO3 +: 390.2727;実測値 390.2749。 1 H NMR (CDCl 3 ): δ = 7.234 (1H, s), 4.298 (2H, t, J = 7.16 Hz), 4.231 (2H, t, J = 6.12 Hz), 3.693 (2H, t, J = 6.08 Hz), 2.698 (2H, t, J = 7.74 Hz), 2.311 (2H, t, J = 7.44 Hz), 2.138 (1H, brs), 1.872 (4H, m), 1.621 (4H, m), 1.327- 1.238 (14H, m), 0.873 (3H, t, J = 6.94 Hz).
13 C NMR (CDCl 3 ): δ = 174.16, 148.50, 120.50, 61.37, 59.08, 50.12, 34.24, 31.82 (× 2), 30.26, 29.54, 29.28, 29.10, 28.84, 28.59, 26.26, 25.72, 24.80, 22.70, 14.16.
HRMS (ESI, [M + Na] + ): Calculated C 20 H 37 N 3 NaO 3 + : 390.2727; Found 390.2749.
13C NMR (CDCl3) :δ= 174.16, 148.50, 120.50, 61.37, 59.08, 50.12, 34.24, 31.82 (×2), 30.26, 29.54, 29.28, 29.10, 28.84, 28.59, 26.26, 25.72, 24.80, 22.70, 14.16。
HRMS (ESI, [M+Na]+):計算値 C20H37N3NaO3 +: 390.2727;実測値 390.2749。 1 H NMR (CDCl 3 ): δ = 7.234 (1H, s), 4.298 (2H, t, J = 7.16 Hz), 4.231 (2H, t, J = 6.12 Hz), 3.693 (2H, t, J = 6.08 Hz), 2.698 (2H, t, J = 7.74 Hz), 2.311 (2H, t, J = 7.44 Hz), 2.138 (1H, brs), 1.872 (4H, m), 1.621 (4H, m), 1.327- 1.238 (14H, m), 0.873 (3H, t, J = 6.94 Hz).
13 C NMR (CDCl 3 ): δ = 174.16, 148.50, 120.50, 61.37, 59.08, 50.12, 34.24, 31.82 (× 2), 30.26, 29.54, 29.28, 29.10, 28.84, 28.59, 26.26, 25.72, 24.80, 22.70, 14.16.
HRMS (ESI, [M + Na] + ): Calculated C 20 H 37 N 3 NaO 3 + : 390.2727; Found 390.2749.
上記製造例と同様の手法により以下の化合物を調製し、実施例14の化合物の製造に使用した。
The following compounds were prepared by the same method as in the above Production Example and used for the production of the compound of Example 14.
[製造例6]3-{8-(4-ヘキシル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}-1-プロパノール
[Production Example 6] 3- {8- (4-Hexyl-1,2,3-triazol-1-yl) octanoyloxy} -1-propanol
1H NMR (CDCl3) :δ=7.260 (1H, s), 4.302 (2H, t, J=7.20 Hz), 2.705 (2H, t, J=7.60 Hz), 2.371 (2H, m), 1.883 (2H, m), 1.641 (4H, m), 1.339-1.276 (12H, m), 0.878 (3H, t, J=7.00 Hz)。
HRMS (MALDI-TOF, [M+Na]-):計算値 C16H29N3NaO2 -: 318.2152;実測値 318.2183。
融点:60.7-61.6℃
実施例15の製造において、以下の化合物を合成中間体として使用した。 1 H NMR (CDCl 3 ): δ = 7.260 (1H, s), 4.302 (2H, t, J = 7.20 Hz), 2.705 (2H, t, J = 7.60 Hz), 2.371 (2H, m), 1.883 ( 2H, m), 1.641 (4H, m), 1.339-1.276 (12H, m), 0.878 (3H, t, J = 7.00 Hz).
HRMS (MALDI-TOF, [M + Na] − ): calculated C 16 H 29 N 3 NaO 2 − : 318.2152; found 318.2183.
Melting point: 60.7-61.6 ℃
In the preparation of Example 15, the following compounds were used as synthetic intermediates.
HRMS (MALDI-TOF, [M+Na]-):計算値 C16H29N3NaO2 -: 318.2152;実測値 318.2183。
融点:60.7-61.6℃
実施例15の製造において、以下の化合物を合成中間体として使用した。 1 H NMR (CDCl 3 ): δ = 7.260 (1H, s), 4.302 (2H, t, J = 7.20 Hz), 2.705 (2H, t, J = 7.60 Hz), 2.371 (2H, m), 1.883 ( 2H, m), 1.641 (4H, m), 1.339-1.276 (12H, m), 0.878 (3H, t, J = 7.00 Hz).
HRMS (MALDI-TOF, [M + Na] − ): calculated C 16 H 29 N 3 NaO 2 − : 318.2152; found 318.2183.
Melting point: 60.7-61.6 ℃
In the preparation of Example 15, the following compounds were used as synthetic intermediates.
[製造例7]3-[6-{4-ヘキシル-(2Z)-2-ブテン-1-イルオキシ}ヘキサノイルオキシ]プロパノール
[Production Example 7] 3- [6- {4-hexyl- (2Z) -2-buten-1-yloxy} hexanoyloxy] propanol
DMF(10ml)中にNaH(968.7mg、24.22mmol)を懸濁させ、0℃に冷却した。懸濁液にDMF(20ml)中のcis-2-ブテン-1,4-ジオール(4.261g、48.36mmol)の溶液を滴下して加え、0℃で1時間攪拌し、その後THF(20ml)中の1-ブロモヘキサン(2.645g、16.02mmol)の溶液を滴下して加え、室温で13時間攪拌した。反応混合物をAcOEt(20ml)およびH2O(30ml)で希釈し、AcOEt(20ml×3)で抽出した。合わせた有機層を水および食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=6:1→4:1)により精製し、表題の化合物を得た(2.320g、13.47mmol、84%、無色の油状物)。
NaH (968.7 mg, 24.22 mmol) was suspended in DMF (10 ml) and cooled to 0 ° C. To the suspension was added dropwise a solution of cis-2-butene-1,4-diol (4.261 g, 48.36 mmol) in DMF (20 ml) and stirred at 0 ° C. for 1 hour, then THF (20 ml The solution of 1-bromohexane (2.645 g, 16.02 mmol) in) was added dropwise and stirred at room temperature for 13 hours. The reaction mixture was diluted with AcOEt (20 ml) and H 2 O (30 ml) and extracted with AcOEt (20 ml × 3). The combined organic layers were washed with water and brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 6: 1 → 4: 1) to give the title compound (2.320 g, 13.47 mmol, 84%, colorless oil).
1H NMR (CDCl3) :δ=5.854-5.787 (1H, m), 5.744-5.682 (1H, m), 4.204 (2H, t, J=5.88 Hz), 4.042 (2H, d, J=6.04 Hz), 3.436 (2H, t, J=6.70 Hz), 2.032 (1H, t, J= 5.90 Hz), 1.582 (2H, quintet, J=6.72 Hz), 1.375-1.251 (6H, m), 0.883 (3H, t, J=6.86 Hz)。
HRMS (ESI, [M+Na]+):計算値 C10H20NaO2 +: 195.1356;実測値:195.1363。
工程2:6-{4-ヘキシル-(2Z)-2-ブテン-1-イルオキシ}ヘキサン酸 1 H NMR (CDCl 3 ): δ = 5.854-5.787 (1H, m), 5.744-5.682 (1H, m), 4.204 (2H, t, J = 5.88 Hz), 4.042 (2H, d, J = 6.04 Hz ), 3.436 (2H, t, J = 6.70 Hz), 2.032 (1H, t, J = 5.90 Hz), 1.582 (2H, quintet, J = 6.72 Hz), 1.375-1.251 (6H, m), 0.883 (3H , t, J = 6.86 Hz).
HRMS (ESI, [M + Na] + ): Calculated C 10 H 20 NaO 2 + : 195.1356; Found: 195.1363.
Step 2: 6- {4-Hexyl- (2Z) -2-buten-1-yloxy} hexanoic acid
HRMS (ESI, [M+Na]+):計算値 C10H20NaO2 +: 195.1356;実測値:195.1363。
工程2:6-{4-ヘキシル-(2Z)-2-ブテン-1-イルオキシ}ヘキサン酸 1 H NMR (CDCl 3 ): δ = 5.854-5.787 (1H, m), 5.744-5.682 (1H, m), 4.204 (2H, t, J = 5.88 Hz), 4.042 (2H, d, J = 6.04 Hz ), 3.436 (2H, t, J = 6.70 Hz), 2.032 (1H, t, J = 5.90 Hz), 1.582 (2H, quintet, J = 6.72 Hz), 1.375-1.251 (6H, m), 0.883 (3H , t, J = 6.86 Hz).
HRMS (ESI, [M + Na] + ): Calculated C 10 H 20 NaO 2 + : 195.1356; Found: 195.1363.
Step 2: 6- {4-Hexyl- (2Z) -2-buten-1-yloxy} hexanoic acid
DMF(10ml)中にNaH(475.8mg、11.90mmol)およびテトラブチルアンモニウムヨージド(654.3mg、1.771mmol)を懸濁させ、0℃で撹拌した。懸濁液にDMF(10ml)中の4-ヘキシルオキシ-(2Z)-2-ブテン-1-オール(1.004g、5.830mmol)の溶液を滴下して加え、0℃で1時間攪拌した。その後DMF(20ml)中の6-ブロモヘキサン酸メチルエステル(1.823g、8.721mmol)の溶液を滴下して加え、50℃で44時間攪拌した。反応混合物をAcOEt(30ml)およびH2O(30ml)で希釈し、AcOEt(30ml×3)で抽出した。合わせた有機層を水および食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=29:1→9:1)により精製し、副生成物を含有する目的のエステルを得た(486.0mg、無色の油状物)。また出発物質のアルコールの一部(313.3mg)も回収した。
NaH (475.8 mg, 11.90 mmol) and tetrabutylammonium iodide (654.3 mg, 1.771 mmol) were suspended in DMF (10 ml) and stirred at 0 ° C. To the suspension was added dropwise a solution of 4-hexyloxy- (2Z) -2-buten-1-ol (1.004 g, 5.830 mmol) in DMF (10 ml) and stirred at 0 ° C. for 1 hour. . Then a solution of 6-bromohexanoic acid methyl ester (1.823 g, 8.721 mmol) in DMF (20 ml) was added dropwise and stirred at 50 ° C. for 44 hours. The reaction mixture was diluted with AcOEt (30 ml) and H 2 O (30 ml) and extracted with AcOEt (30 ml × 3). The combined organic layers were washed with water and brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 29: 1 → 9: 1) to give the desired ester containing by-products (486.0 mg, colorless oil). A portion of the starting alcohol (313.3 mg) was also recovered.
得られた生成物および副生生物の混合物(483.1mg)をメタノール(10ml)に溶解させ、NaOH水溶液(2N、10ml)を加え、室温で3.5時間攪拌した。反応混合物にジエチルエーテル(30ml)を加え、HCl水溶液(1N)を加えて水層のpHを2にして、ジエチルエーテル(30ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=9:1→2:1)により精製し、表題の化合物を得た(358.2mg、1.251mmol、21%、無色の油状物)。
The obtained product and by-product mixture (483.1 mg) was dissolved in methanol (10 ml), aqueous NaOH solution (2N, 10 ml) was added, and the mixture was stirred at room temperature for 3.5 hr. Diethyl ether (30 ml) was added to the reaction mixture, an aqueous HCl solution (1N) was added to adjust the pH of the aqueous layer to 2, and the mixture was extracted with diethyl ether (30 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 9: 1 → 2: 1) to give the title compound (358.2 mg, 1.251 mmol, 21%, colorless oil).
1H NMR (CDCl3) :δ=5.707 (2H, m), 4.033 (4H, d, J=4.60 Hz), 3.436-3.394 (4H, m), 2.360 (2H, t, J=7.48 Hz), 1.689-1.533 (6H, m), 1.421 (2H, m), 1.293 (6H, m), 0.883 (3H, t, J=6.88 Hz)。
HRMS (ESI, [M-H]-):計算値 C16H29O4 -: 285.2071;実測値 285.2070.
工程3:3-[6-{4-ヘキシル-(2Z)-2-ブテン-1-イルオキシ}ヘキサノイルオキシ]プロパノール 1 H NMR (CDCl 3 ): δ = 5.707 (2H, m), 4.033 (4H, d, J = 4.60 Hz), 3.436-3.394 (4H, m), 2.360 (2H, t, J = 7.48 Hz), 1.689-1.533 (6H, m), 1.421 (2H, m), 1.293 (6H, m), 0.883 (3H, t, J = 6.88 Hz).
HRMS (ESI, [MH] - ): Calculated C 16 H 29 O 4 -: 285.2071; Found 285.2070.
Step 3: 3- [6- {4-Hexyl- (2Z) -2-buten-1-yloxy} hexanoyloxy] propanol
HRMS (ESI, [M-H]-):計算値 C16H29O4 -: 285.2071;実測値 285.2070.
工程3:3-[6-{4-ヘキシル-(2Z)-2-ブテン-1-イルオキシ}ヘキサノイルオキシ]プロパノール 1 H NMR (CDCl 3 ): δ = 5.707 (2H, m), 4.033 (4H, d, J = 4.60 Hz), 3.436-3.394 (4H, m), 2.360 (2H, t, J = 7.48 Hz), 1.689-1.533 (6H, m), 1.421 (2H, m), 1.293 (6H, m), 0.883 (3H, t, J = 6.88 Hz).
HRMS (ESI, [MH] - ): Calculated C 16 H 29 O 4 -: 285.2071; Found 285.2070.
Step 3: 3- [6- {4-Hexyl- (2Z) -2-buten-1-yloxy} hexanoyloxy] propanol
1,3-プロパンジオール(324.4mg、4.263mmol)、6-{4-ヘキシル-(2Z)-2-ブテン-1-イルオキシ}ヘキサン酸(300.2mg、1.048mmol)および4-ジメチルアミノピリジン(14.3mg、0.119mmol)をCH2Cl2(10ml)に溶解させ、1-エチル-3-(3-ジメチルアミノプロピル)カルボジイミド塩酸塩(212.5mg、1.109mmol)を0℃で加え、室温で19時間攪拌した。5%KHSO4水溶液(20ml)で反応をクエンチし、有機層を5% KHSO4水溶液(15ml×3)および食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=4:1)により精製し、表題の化合物を得た(315.1mg、0.915mmol、87%、無色の油状物)。
1,3-propanediol (324.4 mg, 4.263 mmol), 6- {4-hexyl- (2Z) -2-buten-1-yloxy} hexanoic acid (300.2 mg, 1.048 mmol) and 4-dimethyl Aminopyridine (14.3 mg, 0.119 mmol) was dissolved in CH 2 Cl 2 (10 ml) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (212.5 mg, 1.109 mmol) was added to 0. The mixture was added at ° C and stirred at room temperature for 19 hours. The reaction was quenched with 5% aqueous KHSO 4 (20 ml) and the organic layer was washed with 5% aqueous KHSO 4 (15 ml × 3) and brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 4: 1) to give the title compound (315.1 mg, 0.915 mmol, 87%, colorless oil).
1H NMR (CDCl3) :δ=5.701 (2H, m), 4.232 (2H, t, J=6.2 Hz), 4.026 (4H, d, J=4.4 Hz), 3.682 (2H, t, J=5.8 Hz), 3.407 (4H, dt, J=6.4, 2.4 Hz), 2.323 (2H, t, J=7.4 Hz), 1.959 (1H, brs), 1.862 (2H, quintet, J=7.4 Hz), 1.684-1.528 (6H, m), 1.422-1.246 (8H, m), 0.880 (3H, t, J=6.8 Hz)。
13C NMR (CDCl3) :δ=174.26, 129.68, 129.45, 70.77, 70.29, 66.65, 66.58, 61.31, 59.30, 34.35, 31.87, 31.83, 29.86, 29.54, 26.00, 25.92, 24.94, 22.76, 14.19。
HRMS (ESI, [M+Na]+):計算値 C19H36NaO5 +: 367.2455;実測値 367.2444。 1 H NMR (CDCl 3 ): δ = 5.701 (2H, m), 4.232 (2H, t, J = 6.2 Hz), 4.026 (4H, d, J = 4.4 Hz), 3.682 (2H, t, J = 5.8 Hz), 3.407 (4H, dt, J = 6.4, 2.4 Hz), 2.323 (2H, t, J = 7.4 Hz), 1.959 (1H, brs), 1.862 (2H, quintet, J = 7.4 Hz), 1.684- 1.528 (6H, m), 1.422-1.246 (8H, m), 0.880 (3H, t, J = 6.8 Hz).
13 C NMR (CDCl 3 ): δ = 174.26, 129.68, 129.45, 70.77, 70.29, 66.65, 66.58, 61.31, 59.30, 34.35, 31.87, 31.83, 29.86, 29.54, 26.00, 25.92, 24.94, 22.76, 14.19.
HRMS (ESI, [M + Na] + ): Calculated C 19 H 36 NaO 5 + : 367.2455; found 367.2444.
13C NMR (CDCl3) :δ=174.26, 129.68, 129.45, 70.77, 70.29, 66.65, 66.58, 61.31, 59.30, 34.35, 31.87, 31.83, 29.86, 29.54, 26.00, 25.92, 24.94, 22.76, 14.19。
HRMS (ESI, [M+Na]+):計算値 C19H36NaO5 +: 367.2455;実測値 367.2444。 1 H NMR (CDCl 3 ): δ = 5.701 (2H, m), 4.232 (2H, t, J = 6.2 Hz), 4.026 (4H, d, J = 4.4 Hz), 3.682 (2H, t, J = 5.8 Hz), 3.407 (4H, dt, J = 6.4, 2.4 Hz), 2.323 (2H, t, J = 7.4 Hz), 1.959 (1H, brs), 1.862 (2H, quintet, J = 7.4 Hz), 1.684- 1.528 (6H, m), 1.422-1.246 (8H, m), 0.880 (3H, t, J = 6.8 Hz).
13 C NMR (CDCl 3 ): δ = 174.26, 129.68, 129.45, 70.77, 70.29, 66.65, 66.58, 61.31, 59.30, 34.35, 31.87, 31.83, 29.86, 29.54, 26.00, 25.92, 24.94, 22.76, 14.19.
HRMS (ESI, [M + Na] + ): Calculated C 19 H 36 NaO 5 + : 367.2455; found 367.2444.
実施例16の製造において、以下の化合物を合成中間体として使用した。
In the production of Example 16, the following compounds were used as synthetic intermediates.
[製造例8]3-ヒドロキシプロピオン酸オレイルエステル
[Production Example 8] 3-Hydroxypropionic acid oleyl ester
工程1:3-tert-ブチルジメチルシリルオキシ-1-プロパノール
TBSO-(CH2)3-OH
NaH(790.9mg、19.77mmol)をTHF(3ml)中に懸濁させ、0℃で攪拌した。懸濁液にTHF(3ml)中の1,3-プロパンジオール(1000.4mg、13.15mmol)の溶液を滴下して加え、0℃で30分間攪拌した。THF(4ml)中のtert-ブチルジメチルシリルクロリド(2.3841g、5.82mmol)の溶液を滴下して加え、室温で22.5時間攪拌した。反応混合物をH2O(20ml)で希釈し、AcOEt(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=8:1)により精製し、表題の化合物を得た(1.9075g、10.02mmol、76%、無色の油状物)。 Step 1: 3-tert-butyldimethylsilyloxy-1-propanol TBSO- (CH 2 ) 3 —OH
NaH (790.9 mg, 19.77 mmol) was suspended in THF (3 ml) and stirred at 0 ° C. To the suspension was added dropwise a solution of 1,3-propanediol (1000.4 mg, 13.15 mmol) in THF (3 ml) and stirred at 0 ° C. for 30 minutes. A solution of tert-butyldimethylsilyl chloride (2.3841 g, 5.82 mmol) in THF (4 ml) was added dropwise and stirred at room temperature for 22.5 hours. The reaction mixture was diluted with H 2 O (20 ml) and extracted with AcOEt (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 8: 1) to give the title compound (1.99075 g, 10.02 mmol, 76%, colorless oil).
TBSO-(CH2)3-OH
NaH(790.9mg、19.77mmol)をTHF(3ml)中に懸濁させ、0℃で攪拌した。懸濁液にTHF(3ml)中の1,3-プロパンジオール(1000.4mg、13.15mmol)の溶液を滴下して加え、0℃で30分間攪拌した。THF(4ml)中のtert-ブチルジメチルシリルクロリド(2.3841g、5.82mmol)の溶液を滴下して加え、室温で22.5時間攪拌した。反応混合物をH2O(20ml)で希釈し、AcOEt(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=8:1)により精製し、表題の化合物を得た(1.9075g、10.02mmol、76%、無色の油状物)。 Step 1: 3-tert-butyldimethylsilyloxy-1-propanol TBSO- (CH 2 ) 3 —OH
NaH (790.9 mg, 19.77 mmol) was suspended in THF (3 ml) and stirred at 0 ° C. To the suspension was added dropwise a solution of 1,3-propanediol (1000.4 mg, 13.15 mmol) in THF (3 ml) and stirred at 0 ° C. for 30 minutes. A solution of tert-butyldimethylsilyl chloride (2.3841 g, 5.82 mmol) in THF (4 ml) was added dropwise and stirred at room temperature for 22.5 hours. The reaction mixture was diluted with H 2 O (20 ml) and extracted with AcOEt (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 8: 1) to give the title compound (1.99075 g, 10.02 mmol, 76%, colorless oil).
1H-NMR(CDCl3):δ= 3.837 (2H, t, J=5.56 Hz), 3.805 (2H, t, J=5.44 Hz), 2.567 (1H, brs), 1.779 (2H, m), 0.900 (9H, s), 0.077 (6H, s)。
工程2:3-tert-ブチルジメチルシリルオキシ-1-プロパナール
TBSO-(CH2)2-CHO
3-tert-ブチルジメチルシリルオキシ-1-プロパノール(1.7995g、9.453mmol)を無水CH2Cl2(10ml)、DMSO(10ml)およびEt3N(6.57ml、47.28mmol)に溶解させ、室温で攪拌しながらSO3-ピリジン錯体(7.5257g、47.28mmol)を加え、室温で1時間攪拌した。反応混合物をEt2O(30ml)およびH2O(20ml)で希釈し、Et2O(20ml×3)で抽出した。合わせた有機層をNa2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=20:1)により精製し、表題の化合物を得た(1.0523g、5.587mmol、59%、無色の油状物)。 1 H-NMR (CDCl 3 ): δ = 3.837 (2H, t, J = 5.56 Hz), 3.805 (2H, t, J = 5.44 Hz), 2.567 (1H, brs), 1.779 (2H, m), 0.900 (9H, s), 0.077 (6H, s).
Step 2: 3-tert-butyldimethylsilyloxy-1-propanal TBSO— (CH 2 ) 2 —CHO
3-tert-butyldimethylsilyloxy-1-propanol (1.7955 g, 9.453 mmol) was dissolved in anhydrous CH 2 Cl 2 (10 ml), DMSO (10 ml) and Et 3 N (6.57 ml, 47.28 mmol). SO 3 -pyridine complex (7.5257 g, 47.28 mmol) was added with stirring at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with Et 2 O (30 ml) and H 2 O (20 ml) and extracted with Et 2 O (20 ml × 3). The combined organic layers were dried over Na 2 SO 4 and the solvent was distilled off. The residue was purified by column chromatography (hexane / AcOEt = 20: 1) to give the title compound (1.0523 g, 5.587 mmol, 59%, colorless oil).
工程2:3-tert-ブチルジメチルシリルオキシ-1-プロパナール
TBSO-(CH2)2-CHO
3-tert-ブチルジメチルシリルオキシ-1-プロパノール(1.7995g、9.453mmol)を無水CH2Cl2(10ml)、DMSO(10ml)およびEt3N(6.57ml、47.28mmol)に溶解させ、室温で攪拌しながらSO3-ピリジン錯体(7.5257g、47.28mmol)を加え、室温で1時間攪拌した。反応混合物をEt2O(30ml)およびH2O(20ml)で希釈し、Et2O(20ml×3)で抽出した。合わせた有機層をNa2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=20:1)により精製し、表題の化合物を得た(1.0523g、5.587mmol、59%、無色の油状物)。 1 H-NMR (CDCl 3 ): δ = 3.837 (2H, t, J = 5.56 Hz), 3.805 (2H, t, J = 5.44 Hz), 2.567 (1H, brs), 1.779 (2H, m), 0.900 (9H, s), 0.077 (6H, s).
Step 2: 3-tert-butyldimethylsilyloxy-1-propanal TBSO— (CH 2 ) 2 —CHO
3-tert-butyldimethylsilyloxy-1-propanol (1.7955 g, 9.453 mmol) was dissolved in anhydrous CH 2 Cl 2 (10 ml), DMSO (10 ml) and Et 3 N (6.57 ml, 47.28 mmol). SO 3 -pyridine complex (7.5257 g, 47.28 mmol) was added with stirring at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with Et 2 O (30 ml) and H 2 O (20 ml) and extracted with Et 2 O (20 ml × 3). The combined organic layers were dried over Na 2 SO 4 and the solvent was distilled off. The residue was purified by column chromatography (hexane / AcOEt = 20: 1) to give the title compound (1.0523 g, 5.587 mmol, 59%, colorless oil).
1H-NMR(CDCl3):δ=9.802 (1H, t, J=2.04 Hz), 3.986 (2H, t, J=6.02 Hz), 2.596 (2H, dt, J=6.04, 2.04 Hz), 0.880 (9H, s), 0.064 (6H, s)。
工程3:3-tert-ブチルジメチルシリルオキシ-プロピオン酸
TBSO-(CH2)2-COOH
3-tert-ブチルジメチルシリルオキシ-1-プロパナール(1.0438g、5.542mmol)および2-メチル-2-ブテン(1.7191g、24.51mmol)をtBuOH (15ml)に溶解させ、H2O(5ml)中のNaH2PO4・2H2O(867.8mg、5.562mmol)およびNaClO2(2.0425g、22.58mmol)の溶液を加え、室温で1時間攪拌した。tBuOHを留去し、残渣に5%KHSO4水溶液を加え、AcOEt(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=2:1)により精製し、表題の化合物を得た(946.5mg、4.632mmol、84%、無色の油状物)。 1 H-NMR (CDCl 3 ): δ = 9.802 (1H, t, J = 2.04 Hz), 3.986 (2H, t, J = 6.02 Hz), 2.596 (2H, dt, J = 6.04, 2.04 Hz), 0.880 (9H, s), 0.064 (6H, s).
Step 3: 3-tert-butyldimethylsilyloxy-propionic acid TBSO- (CH 2 ) 2 —COOH
3-tert-butyldimethylsilyloxy-1-propanal (1.0438 g, 5.542 mmol) and 2-methyl-2-butene (1.7191 g, 24.51 mmol) were dissolved in t BuOH (15 ml) and H A solution of NaH 2 PO 4 .2H 2 O (867.8 mg, 5.562 mmol) and NaClO 2 (2.0425 g, 22.58 mmol) in 2 O (5 ml) was added and stirred at room temperature for 1 hour. t BuOH was distilled off, 5% KHSO 4 aqueous solution was added to the residue, and the mixture was extracted with AcOEt (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 2: 1) to give the title compound (946.5 mg, 4.632 mmol, 84%, colorless oil).
工程3:3-tert-ブチルジメチルシリルオキシ-プロピオン酸
TBSO-(CH2)2-COOH
3-tert-ブチルジメチルシリルオキシ-1-プロパナール(1.0438g、5.542mmol)および2-メチル-2-ブテン(1.7191g、24.51mmol)をtBuOH (15ml)に溶解させ、H2O(5ml)中のNaH2PO4・2H2O(867.8mg、5.562mmol)およびNaClO2(2.0425g、22.58mmol)の溶液を加え、室温で1時間攪拌した。tBuOHを留去し、残渣に5%KHSO4水溶液を加え、AcOEt(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=2:1)により精製し、表題の化合物を得た(946.5mg、4.632mmol、84%、無色の油状物)。 1 H-NMR (CDCl 3 ): δ = 9.802 (1H, t, J = 2.04 Hz), 3.986 (2H, t, J = 6.02 Hz), 2.596 (2H, dt, J = 6.04, 2.04 Hz), 0.880 (9H, s), 0.064 (6H, s).
Step 3: 3-tert-butyldimethylsilyloxy-propionic acid TBSO- (CH 2 ) 2 —COOH
3-tert-butyldimethylsilyloxy-1-propanal (1.0438 g, 5.542 mmol) and 2-methyl-2-butene (1.7191 g, 24.51 mmol) were dissolved in t BuOH (15 ml) and H A solution of NaH 2 PO 4 .2H 2 O (867.8 mg, 5.562 mmol) and NaClO 2 (2.0425 g, 22.58 mmol) in 2 O (5 ml) was added and stirred at room temperature for 1 hour. t BuOH was distilled off, 5% KHSO 4 aqueous solution was added to the residue, and the mixture was extracted with AcOEt (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 2: 1) to give the title compound (946.5 mg, 4.632 mmol, 84%, colorless oil).
1H-NMR(CDCl3):δ=10.548 (1H, brs), 3.918 (2H, t, J=6.14 Hz), 2.585 (2H, t, J=6.18 Hz), 0.895 (9H, s), 0.086 (6H, s)。
1 H-NMR (CDCl 3 ): δ = 10.548 (1H, brs), 3.918 (2H, t, J = 6.14 Hz), 2.585 (2H, t, J = 6.18 Hz), 0.895 (9H, s), 0.086 (6H, s).
工程4:3-tert-ブチルジメチルシリルオキシプロピオン酸オレイルエステル
Process 4: 3-tert-butyldimethylsilyloxypropionic acid oleyl ester
オレイルアルコール(208.1mg、0.775mmol)、3-tert-ブチルジメチルシリルオキシ-プロピオン酸(304.2mg、1.489mmol)および4-ジメチルアミノピリジン(21.6mg、0.180mmol)をCH2Cl2(5ml)に溶解させ、1-エチル-3-(3-ジメチルアミノプロピル)カルボジイミド塩酸塩(341.1mg、1.779mmol)を0℃で加え、室温で19時間攪拌した。H2O(10ml)を加えて反応をクエンチし、CH2Cl2(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=8:1)により精製し、表題の化合物を得た(345.1mg、0.759mmol、98%、無色の油状物)。
Oleyl alcohol (208.1 mg, 0.775 mmol), 3-tert-butyldimethylsilyloxy-propionic acid (304.2 mg, 1.490 mmol) and 4-dimethylaminopyridine (21.6 mg, 0.180 mmol) in CH 2 Dissolved in Cl 2 (5 ml), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (341.1 mg, 1.779 mmol) was added at 0 ° C. and stirred at room temperature for 19 hours. The reaction was quenched with H 2 O (10 ml) and extracted with CH 2 Cl 2 (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 8: 1) to give the title compound (345.1 mg, 0.759 mmol, 98%, colorless oil).
1H-NMR(CDCl3):δ=5.347 (2H, m), 4.070 (2H, t, J=6.76 Hz), 3.895 (2H, t, J=6.42 Hz), 2.514 (2H, t, J=6.42 Hz), 2.012 (4H, m), 1.618 (2H, m), 1.267 (22H, m), 0.878 (12H, m), 0.054 (6H, s)。
1 H-NMR (CDCl 3 ): δ = 5.347 (2H, m), 4.070 (2H, t, J = 6.76 Hz), 3.895 (2H, t, J = 6.42 Hz), 2.514 (2H, t, J = 6.42 Hz), 2.012 (4H, m), 1.618 (2H, m), 1.267 (22H, m), 0.878 (12H, m), 0.054 (6H, s).
工程5:3-ヒドロキシプロピオン酸オレイルエステル
Process 5: 3-Hydroxypropionic acid oleyl ester
THF(5ml)中の3-tert-ブチルジメチルシリルオキシプロピオン酸オレイルエステル(300.1mg、0.660mmol)の溶液にHF・ピリジン錯体(70%、0.120ml)を加え、室温で12時間攪拌した。H2O(10ml)を加え、AcOEt(10ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、要馬を留去した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=4:1)により精製し、表題の化合物を得た(195.7mg、0.575mmol、87%、黄色の油状物)。
HF • pyridine complex (70%, 0.120 ml) was added to a solution of 3-tert-butyldimethylsilyloxypropionic acid oleyl ester (300.1 mg, 0.660 mmol) in THF (5 ml) and stirred at room temperature for 12 hours. did. H 2 O (10 ml) was added and extracted with AcOEt (10 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and the main horse was distilled off. The residue was purified by column chromatography (hexane: AcOEt = 4: 1) to give the title compound (195.7 mg, 0.575 mmol, 87%, yellow oil).
1H-NMR(CDCl3):δ=5.345 (2H, m), 4.109 (2H, t, J=6.60 Hz), 3.865 (2H, t, J=5.20 Hz), 2.571 (2H, t, J=5.60 Hz), 2.416 (1H, brs), 2.011 (4H, m), 1.635 (2H, m), 1.265 (22H, m), 0.878 (3H, t, J=6.68 Hz)。
13C-NMR(CDCl3):δ=173.03, 129.99, 129.76, 64.91, 58.32, 36.70, 31.89, 29.75, 29.71, 29.51, 29.38, 29.31, 29.22, 29.19, 28.55, 27.21, 27.17, 25.87, 22.67, 14.10。
HRMS (ESI, [M+Na]+):計算値 C21H40NaO3 +: 363.2870;実測値:363.2853。 1 H-NMR (CDCl 3 ): δ = 5.345 (2H, m), 4.109 (2H, t, J = 6.60 Hz), 3.865 (2H, t, J = 5.20 Hz), 2.571 (2H, t, J = 5.60 Hz), 2.416 (1H, brs), 2.011 (4H, m), 1.635 (2H, m), 1.265 (22H, m), 0.878 (3H, t, J = 6.68 Hz).
13 C-NMR (CDCl 3 ): δ = 173.03, 129.99, 129.76, 64.91, 58.32, 36.70, 31.89, 29.75, 29.71, 29.51, 29.38, 29.31, 29.22, 29.19, 28.55, 27.21, 27.17, 25.87, 22.67, 14.10 .
HRMS (ESI, [M + Na] + ): Calculated C 21 H 40 NaO 3 + : 363.2870; Found: 363.22853.
13C-NMR(CDCl3):δ=173.03, 129.99, 129.76, 64.91, 58.32, 36.70, 31.89, 29.75, 29.71, 29.51, 29.38, 29.31, 29.22, 29.19, 28.55, 27.21, 27.17, 25.87, 22.67, 14.10。
HRMS (ESI, [M+Na]+):計算値 C21H40NaO3 +: 363.2870;実測値:363.2853。 1 H-NMR (CDCl 3 ): δ = 5.345 (2H, m), 4.109 (2H, t, J = 6.60 Hz), 3.865 (2H, t, J = 5.20 Hz), 2.571 (2H, t, J = 5.60 Hz), 2.416 (1H, brs), 2.011 (4H, m), 1.635 (2H, m), 1.265 (22H, m), 0.878 (3H, t, J = 6.68 Hz).
13 C-NMR (CDCl 3 ): δ = 173.03, 129.99, 129.76, 64.91, 58.32, 36.70, 31.89, 29.75, 29.71, 29.51, 29.38, 29.31, 29.22, 29.19, 28.55, 27.21, 27.17, 25.87, 22.67, 14.10 .
HRMS (ESI, [M + Na] + ): Calculated C 21 H 40 NaO 3 + : 363.2870; Found: 363.22853.
上記製造例と同様の手法により以下の化合物を調製し、実施例17および18の化合物の製造に使用した。
The following compounds were prepared by the same method as in the above production examples and used for the production of the compounds of Examples 17 and 18.
[製造例9]4-ヒドロキシブタン酸オレイルエステル
[Production Example 9] 4-Hydroxybutanoic acid oleyl ester
1H-NMR(CDCl3):δ=5.340 (2H, m), 4.065 (2H, t, J=6.78 Hz), 3.684 (2H, t, J=6.12 Hz), 2.431 (2H, t, J=7.16 Hz), 2.014 (4H, m), 1.881 (2H, m), 1.830 (1H, brs), 1.614 (2H, m), 1.294 (22H, m), 0.874 (3H, t, J=6.74 Hz)。
13C-NMR(CDCl3):δ=174.04, 129.96, 129.76, 64.74, 62.12, 31.88, 31.09, 29.74, 29.71, 29.50, 29.38, 29.30, 29.20, 29.18, 28.58, 27.71, 27.19, 27.16, 25.88, 22.66, 22.55, 14.08。
HRMS (ESI, [M+Na]+):計算値 C22H42NaO3 +:377.3026;実測値:377.3006。
[製造例10]5-ヒドロキシペンタン酸オレイルエステル 1 H-NMR (CDCl 3 ): δ = 5.340 (2H, m), 4.065 (2H, t, J = 6.78 Hz), 3.684 (2H, t, J = 6.12 Hz), 2.431 (2H, t, J = 7.16 Hz), 2.014 (4H, m), 1.881 (2H, m), 1.830 (1H, brs), 1.614 (2H, m), 1.294 (22H, m), 0.874 (3H, t, J = 6.74 Hz) .
13 C-NMR (CDCl 3 ): δ = 174.04, 129.96, 129.76, 64.74, 62.12, 31.88, 31.09, 29.74, 29.71, 29.50, 29.38, 29.30, 29.20, 29.18, 28.58, 27.71, 27.19, 27.16, 25.88, 22.66 , 22.55, 14.08.
HRMS (ESI, [M + Na] + ): calculated C 22 H 42 NaO 3 + : 377.3026; found: 377.3006.
[Production Example 10] 5-Hydroxypentanoic acid oleyl ester
13C-NMR(CDCl3):δ=174.04, 129.96, 129.76, 64.74, 62.12, 31.88, 31.09, 29.74, 29.71, 29.50, 29.38, 29.30, 29.20, 29.18, 28.58, 27.71, 27.19, 27.16, 25.88, 22.66, 22.55, 14.08。
HRMS (ESI, [M+Na]+):計算値 C22H42NaO3 +:377.3026;実測値:377.3006。
[製造例10]5-ヒドロキシペンタン酸オレイルエステル 1 H-NMR (CDCl 3 ): δ = 5.340 (2H, m), 4.065 (2H, t, J = 6.78 Hz), 3.684 (2H, t, J = 6.12 Hz), 2.431 (2H, t, J = 7.16 Hz), 2.014 (4H, m), 1.881 (2H, m), 1.830 (1H, brs), 1.614 (2H, m), 1.294 (22H, m), 0.874 (3H, t, J = 6.74 Hz) .
13 C-NMR (CDCl 3 ): δ = 174.04, 129.96, 129.76, 64.74, 62.12, 31.88, 31.09, 29.74, 29.71, 29.50, 29.38, 29.30, 29.20, 29.18, 28.58, 27.71, 27.19, 27.16, 25.88, 22.66 , 22.55, 14.08.
HRMS (ESI, [M + Na] + ): calculated C 22 H 42 NaO 3 + : 377.3026; found: 377.3006.
[Production Example 10] 5-Hydroxypentanoic acid oleyl ester
1H-NMR(CDCl3):δ=5.345 (2H, m), 4.061 (2H, t, J=6.76 Hz), 3.645 (2H, t, J=6.32 Hz), 2.346 (2H, t, J=7.24 Hz), 2.010 (4H, m), 1.723 (2H, m), 1.609 (4H, m), 1.491 (1H, brs), 1.266 (22H, m), 0.878 (3H, t, J=6.68 Hz)。
13C-NMR(CDCl3):δ=173.81, 129.97, 129.77, 64.57, 62.27, 33.90, 32.08, 31.89, 29.75, 29.72, 29.51, 29.40, 29.31, 29.21, 29.19, 28.62, 27.20, 27.17, 25.90, 22.67, 21.06, 14.10。
HRMS (ESI, [M+Na]+):計算値:C23H44NaO3 +: 391.3183;実測値:391.3173。 1 H-NMR (CDCl 3 ): δ = 5.345 (2H, m), 4.061 (2H, t, J = 6.76 Hz), 3.645 (2H, t, J = 6.32 Hz), 2.346 (2H, t, J = 7.24 Hz), 2.010 (4H, m), 1.723 (2H, m), 1.609 (4H, m), 1.491 (1H, brs), 1.266 (22H, m), 0.878 (3H, t, J = 6.68 Hz) .
13 C-NMR (CDCl 3 ): δ = 173.81, 129.97, 129.77, 64.57, 62.27, 33.90, 32.08, 31.89, 29.75, 29.72, 29.51, 29.40, 29.31, 29.21, 29.19, 28.62, 27.20, 27.17, 25.90, 22.67 , 21.06, 14.10.
HRMS (ESI, [M + Na] + ): Calculated: C 23 H 44 NaO 3 + : 391.3183; Found: 391.3173.
13C-NMR(CDCl3):δ=173.81, 129.97, 129.77, 64.57, 62.27, 33.90, 32.08, 31.89, 29.75, 29.72, 29.51, 29.40, 29.31, 29.21, 29.19, 28.62, 27.20, 27.17, 25.90, 22.67, 21.06, 14.10。
HRMS (ESI, [M+Na]+):計算値:C23H44NaO3 +: 391.3183;実測値:391.3173。 1 H-NMR (CDCl 3 ): δ = 5.345 (2H, m), 4.061 (2H, t, J = 6.76 Hz), 3.645 (2H, t, J = 6.32 Hz), 2.346 (2H, t, J = 7.24 Hz), 2.010 (4H, m), 1.723 (2H, m), 1.609 (4H, m), 1.491 (1H, brs), 1.266 (22H, m), 0.878 (3H, t, J = 6.68 Hz) .
13 C-NMR (CDCl 3 ): δ = 173.81, 129.97, 129.77, 64.57, 62.27, 33.90, 32.08, 31.89, 29.75, 29.72, 29.51, 29.40, 29.31, 29.21, 29.19, 28.62, 27.20, 27.17, 25.90, 22.67 , 21.06, 14.10.
HRMS (ESI, [M + Na] + ): Calculated: C 23 H 44 NaO 3 + : 391.3183; Found: 391.3173.
実施例19~23の化合物は、以下の化合物を合成中間体として使用して製造した。
The compounds of Examples 19 to 23 were produced using the following compounds as synthetic intermediates.
[製造例11](2S,3S)-3-[(ジイソプロピルアミノ)-tert-ブトキシホスフィノオキシ]-2-(tert-ブトキシカルボニルアミノ)-3-メチル-プロピオン酸tert-ブチルエステル
[Production Example 11] (2S, 3S) -3-[(diisopropylamino) -tert-butoxyphosphinooxy] -2- (tert-butoxycarbonylamino) -3-methyl-propionic acid tert-butyl ester
工程1:N-tert-ブトキシカルボニル-L-アロ-スレオニンtert-ブチルエステルの調製
Step 1: Preparation of N-tert-butoxycarbonyl-L-allo-threonine tert-butyl ester
L-アロ-スレオニン(997.3mg、8.372mmol)をNaOH水溶液(1N、10ml)、H2O(10ml)およびジオキサン(20ml)の混合液に溶解させ、0℃で攪拌し、ジ-tert-ブチルジカルボネート(2.763g、12.66mmol)を10分かけて加え、室温で24時間攪拌した。反応混合物に5%KHSO4水溶液を加えてpHを3とし、AcOEt(40ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去し、目的のN-Boc-L-アロ-スレオニンを得た(2.016g、無色の油状物)。
L-Allo-threonine (997.3 mg, 8.372 mmol) was dissolved in a mixture of aqueous NaOH (1N, 10 ml), H 2 O (10 ml) and dioxane (20 ml), stirred at 0 ° C. and di-tert. -Butyl dicarbonate (2.763 g, 12.66 mmol) was added over 10 minutes and stirred at room temperature for 24 hours. The reaction mixture was added with 5% KHSO 4 aqueous solution to adjust the pH to 3, and extracted with AcOEt (40 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated to give the desired N-Boc-L-allo-threonine (2.016 g, colorless oil).
N-Boc-L-アロ-スレオニン(2.105g)を無水ジクロロメタン(150ml)に溶解させ、2-tert-ブチル-1,3-ジイソプロピル-イソウレア(5.872g、29.31mmol)を加え、室温で18時間攪拌した。反応混合物にヘキサンを加え、10分間攪拌し、セライト(登録商標)で濾過し、濾液を濃縮した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=4:1)により精製して、表題の化合物を得た(1.581g、5.741mmol、69%、白色固体)。
N-Boc-L-Allo-threonine (2.105 g) was dissolved in anhydrous dichloromethane (150 ml), 2-tert-butyl-1,3-diisopropyl-isourea (5.872 g, 29.31 mmol) was added, and room temperature was added. For 18 hours. Hexane was added to the reaction mixture, stirred for 10 minutes, filtered through Celite (registered trademark), and the filtrate was concentrated. The residue was purified by column chromatography (hexane: AcOEt = 4: 1) to give the title compound (1.581 g, 5.741 mmol, 69%, white solid).
1H-NMR(CDCl3):δ=5.226 (1H, m), 4.224 (1H, s), 4.123 (1H, m), 1.960 (1H, d, J=6.00 Hz), 1.486 (9H, s), 1.455 (9H, s), 1.234 (3H, d, J=6.40 Hz)。
HRMS (ESI, [M+Na]+):計算値:C13H25NNaO5 +: 298.1630;実測値:298.1609。
融点:62.1-63.0 ℃。 1 H-NMR (CDCl 3 ): δ = 5.226 (1H, m), 4.224 (1H, s), 4.123 (1H, m), 1.960 (1H, d, J = 6.00 Hz), 1.486 (9H, s) , 1.455 (9H, s), 1.234 (3H, d, J = 6.40 Hz).
HRMS (ESI, [M + Na] + ): Calculated: C 13 H 25 NNaO 5 + : 298.1630; Found: 298.1609.
Melting point: 62.1-63.0 ° C.
HRMS (ESI, [M+Na]+):計算値:C13H25NNaO5 +: 298.1630;実測値:298.1609。
融点:62.1-63.0 ℃。 1 H-NMR (CDCl 3 ): δ = 5.226 (1H, m), 4.224 (1H, s), 4.123 (1H, m), 1.960 (1H, d, J = 6.00 Hz), 1.486 (9H, s) , 1.455 (9H, s), 1.234 (3H, d, J = 6.40 Hz).
HRMS (ESI, [M + Na] + ): Calculated: C 13 H 25 NNaO 5 + : 298.1630; Found: 298.1609.
Melting point: 62.1-63.0 ° C.
工程2:(2S,3S)-3-[(ジイソプロピルアミノ)-tert-ブトキシホスフィノオキシ]-2-(tert-ブトキシカルボニルアミノ)-3-メチル-プロピオン酸tert-ブチルエステル
Step 2: (2S, 3S) -3-[(Diisopropylamino) -tert-butoxyphosphinooxy] -2- (tert-butoxycarbonylamino) -3-methyl-propionic acid tert-butyl ester
ビス(ジイソプロピルアミノ)-tert-ブチルホスフィン(303.1mg、1.101mmol)をCH2Cl2(5ml)およびトルエン(0.5ml)に溶解させ、N-tert-ブトキシカルボニル-L-アロ-スレオニンtert-ブチルエステル(508.2mg、1.669mmol)を加え、減圧下溶媒を留去した。残渣にCH2Cl2(5ml)を加え、THF(5ml)中の1H-テトラゾール(105.9mg、1.512mmol)の溶液を室温で加えた。数分後に白色固体が析出した。飽和NaHCO3水溶液(10ml)を加えて反応をクエンチし、CH2Cl2(10ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt:Et3N=35:4:1)により精製し、表題の化合物を得た(445.0mg、0.930mmol、84%、黄色油状物)。なお、カラムクロマトグラフィーは、溶離液に3%(v/v)Et3Nを加えてシリカゲルを不活性化して行った。
Bis (diisopropylamino) -tert-butylphosphine (303.1 mg, 1.101 mmol) was dissolved in CH 2 Cl 2 (5 ml) and toluene (0.5 ml), and N-tert-butoxycarbonyl-L-allo-threonine was dissolved. tert-Butyl ester (508.2 mg, 1.669 mmol) was added, and the solvent was distilled off under reduced pressure. To the residue was added CH 2 Cl 2 (5 ml) and a solution of 1H-tetrazole (105.9 mg, 1.512 mmol) in THF (5 ml) was added at room temperature. A white solid precipitated after a few minutes. The reaction was quenched by the addition of saturated aqueous NaHCO 3 (10 ml) and extracted with CH 2 Cl 2 (10 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane: AcOEt: Et 3 N = 35: 4: 1) to give the title compound (445.0 mg, 0.930 mmol, 84%, yellow oil). Column chromatography was performed by adding 3% (v / v) Et 3 N to the eluent to inactivate the silica gel.
1H-NMR(CD2Cl2):δ=5.825 (1/2H, m), 5.300 (1/2H, m), 4.085-3.959 (2H, m), 3.576 (2H, m), 1.436-1.431 (9H, m), 1.405-1.396 (9H, m), 1.360-1.308 (9H, m), 1.270 (3H, d, J= 6.56 Hz), 1.143 (12H, m)。
HRMS (ESI, [M+Na]+):計算値:C23H47N2NaO6P+: 501.3064;実測値:501.3053。 1 H-NMR (CD 2 Cl 2 ): δ = 5.825 (1 / 2H, m), 5.300 (1 / 2H, m), 4.085-3.959 (2H, m), 3.576 (2H, m), 1.436-1.431 (9H, m), 1.405-1.396 (9H, m), 1.360-1.308 (9H, m), 1.270 (3H, d, J = 6.56 Hz), 1.143 (12H, m).
HRMS (ESI, [M + Na] + ): Calculated: C 23 H 47 N 2 NaO 6 P + : 501.3064; Found: 501.3305.
HRMS (ESI, [M+Na]+):計算値:C23H47N2NaO6P+: 501.3064;実測値:501.3053。 1 H-NMR (CD 2 Cl 2 ): δ = 5.825 (1 / 2H, m), 5.300 (1 / 2H, m), 4.085-3.959 (2H, m), 3.576 (2H, m), 1.436-1.431 (9H, m), 1.405-1.396 (9H, m), 1.360-1.308 (9H, m), 1.270 (3H, d, J = 6.56 Hz), 1.143 (12H, m).
HRMS (ESI, [M + Na] + ): Calculated: C 23 H 47 N 2 NaO 6 P + : 501.3064; Found: 501.3305.
[実施例24](2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸
[Example 24] (2S, 3S) -2-Amino-3-[((2R) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid
工程1:(R)-4-(4-メトキシフェノキシ)メチル-2,2-ジメチル-1,3-ジオキソランの調製
Step 1: Preparation of (R) -4- (4-methoxyphenoxy) methyl-2,2-dimethyl-1,3-dioxolane
(R)-(-)-2,2-ジメチル-1,3-ジオキソラン-4-メタノール(3.0058g、22.74mmol)、トリフェニルホスフィン(7.7501g、29.55mmol)、およびp-メトキシフェノール(8.4931g、68.42mmol)を無水トルエン(30ml)に溶解させ、ジエチルアゾジカルボキシレート(2Mトルエン溶液、14.8ml、29.6mmol)を加え、70℃で5時間攪拌した。反応混合物を濃縮し、残渣をカラムクロマトグラフィー(Et2O:ヘキサン=1:15)により精製し、表題の化合物を得た(5.1297g、21.53mmol、95%、無色の油状物)。
(R)-(−)-2,2-dimethyl-1,3-dioxolane-4-methanol (3.054 g, 22.74 mmol), triphenylphosphine (7.7501 g, 29.55 mmol), and p-methoxy Phenol (8.4931 g, 68.42 mmol) was dissolved in anhydrous toluene (30 ml), diethyl azodicarboxylate (2M toluene solution, 14.8 ml, 29.6 mmol) was added, and the mixture was stirred at 70 ° C. for 5 hours. The reaction mixture was concentrated and the residue was purified by column chromatography (Et 2 O: hexane = 1: 15) to give the title compound (5.1297 g, 21.53 mmol, 95%, colorless oil).
1H-NMR(CDCl3):δ=6.841 (4H, m), 4.461 (1H, quintet, J=6.00 Hz), 4.158 (1H, m), 4.018 (1H, m), 3.889 (2H, m), 3.769(3H, s), 1.462 (3H, s), 1.402 (3H, s)。
1 H-NMR (CDCl 3 ): δ = 6.841 (4H, m), 4.461 (1H, quintet, J = 6.00 Hz), 4.158 (1H, m), 4.018 (1H, m), 3.889 (2H, m) , 3.769 (3H, s), 1.462 (3H, s), 1.402 (3H, s).
工程2:(2S)-3-(4-メトキシフェノキシ)-1,2-プロパンジオールの調製
Step 2: Preparation of (2S) -3- (4-methoxyphenoxy) -1,2-propanediol
(R)-4-(4-メトキシフェノキシ)メチル-2,2-ジメチル-1,3-ジオキソラン(6.105g、25.62mmol)をMeOH(150ml)に溶解させ、Amberlyst-15(3.959g)を加え、室温で2日間攪拌した。濾過によりAmberlyst-15を除去し、濾液を濃縮した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=1:1)により精製し、表題の化合物を得た(5.073g、25.59mmol、定量的、白色固体)。
(R) -4- (4-Methoxyphenoxy) methyl-2,2-dimethyl-1,3-dioxolane (6.105 g, 25.62 mmol) was dissolved in MeOH (150 ml) and Amberlyst-15 (3.959 g). ) And stirred at room temperature for 2 days. Amberlyst-15 was removed by filtration and the filtrate was concentrated. The residue was purified by column chromatography (hexane: AcOEt = 1: 1) to give the title compound (5.073 g, 25.59 mmol, quantitative, white solid).
1H-NMR(CDCl3):δ=6.848 (4H, m), 4.089 (1H, m), 4.013 (2H, m), 3.830 (1H, m) , 3.774 (3H, s) , 3.752 (1H, m) , 2.572 (1H, d, J=4.72 Hz), 1.979 (1H, t, J=6.08 Hz)。
融点:73.2-74.5℃。
工程3:(2R)-1,2-ビス(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)プロパンの調製 1 H-NMR (CDCl 3 ): δ = 6.848 (4H, m), 4.089 (1H, m), 4.013 (2H, m), 3.830 (1H, m), 3.774 (3H, s), 3.752 (1H, m), 2.572 (1H, d, J = 4.72 Hz), 1.979 (1H, t, J = 6.08 Hz).
Melting point: 73.2-74.5 ° C.
Step 3: Preparation of (2R) -1,2-bis (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) propane
融点:73.2-74.5℃。
工程3:(2R)-1,2-ビス(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)プロパンの調製 1 H-NMR (CDCl 3 ): δ = 6.848 (4H, m), 4.089 (1H, m), 4.013 (2H, m), 3.830 (1H, m), 3.774 (3H, s), 3.752 (1H, m), 2.572 (1H, d, J = 4.72 Hz), 1.979 (1H, t, J = 6.08 Hz).
Melting point: 73.2-74.5 ° C.
Step 3: Preparation of (2R) -1,2-bis (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) propane
(2S)-3-(4-メトキシフェノキシ)-1,2-プロパンジオール(504.8mg、2.55mmol)およびイミダゾール(519.2mg、7.63mmol)を無水DMF(2.2ml)に溶解させ、室温でtert-ブチルジメチルシリルクロリド(1060.8mg、7.04mmol)を加え、69時間攪拌した。反応混合物を水(15ml)およびAcOEt(20ml)で希釈し、水層をAcOEt(30ml×3)で抽出した。合わせた有機層をNa2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=4:1)により精製し、表題の化合物を得た(1.072g、2.51mmol、98%、無色の油状物)。
(2S) -3- (4-Methoxyphenoxy) -1,2-propanediol (504.8 mg, 2.55 mmol) and imidazole (519.2 mg, 7.63 mmol) were dissolved in anhydrous DMF (2.2 ml). Then, tert-butyldimethylsilyl chloride (1060.8 mg, 7.04 mmol) was added at room temperature, and the mixture was stirred for 69 hours. The reaction mixture was diluted with water (15 ml) and AcOEt (20 ml) and the aqueous layer was extracted with AcOEt (30 ml × 3). The combined organic layers were dried over Na 2 SO 4 and the solvent was distilled off. The residue was purified by column chromatography (hexane: AcOEt = 4: 1) to give the title compound (1.072 g, 2.51 mmol, 98%, colorless oil).
1H-NMR(CDCl3):δ=6.828 (4H, m), 4.025 (2H, m), 3.806 (1H, m), 3.767 (3H, s), 3.632 (2H, d, J=5.56 Hz), 0.893 (9H, s), 0.891 (9H, s), 0.104-0.060 (12H)。
HRMS (ESI, [M+Na]+):計算値:C22H42NaO4Si2 +: 449.2514;実測値449.2517。
工程4:(2R)-2-(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)-1-プロパノールの調製 1 H-NMR (CDCl 3 ): δ = 6.828 (4H, m), 4.025 (2H, m), 3.806 (1H, m), 3.767 (3H, s), 3.632 (2H, d, J = 5.56 Hz) , 0.893 (9H, s), 0.891 (9H, s), 0.104-0.060 (12H).
HRMS (ESI, [M + Na] + ): Calculated: C 22 H 42 NaO 4 Si 2 + : 449.2514; found 449.2517.
Step 4: Preparation of (2R) -2- (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) -1-propanol
HRMS (ESI, [M+Na]+):計算値:C22H42NaO4Si2 +: 449.2514;実測値449.2517。
工程4:(2R)-2-(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)-1-プロパノールの調製 1 H-NMR (CDCl 3 ): δ = 6.828 (4H, m), 4.025 (2H, m), 3.806 (1H, m), 3.767 (3H, s), 3.632 (2H, d, J = 5.56 Hz) , 0.893 (9H, s), 0.891 (9H, s), 0.104-0.060 (12H).
HRMS (ESI, [M + Na] + ): Calculated: C 22 H 42 NaO 4 Si 2 + : 449.2514; found 449.2517.
Step 4: Preparation of (2R) -2- (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) -1-propanol
HF・ピリジン錯体(70%、1.2ml)をピリジン(5.5ml)に溶解させ、当該溶液をTHF(20ml)中の(2R)-1,2-ビス(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)プロパン(2.9927g、7.013mmol)の溶液に加え、室温で13時間攪拌した。反応混合物をAcOEt(20ml)で希釈し、食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=4:1)により精製し、表題の化合物を得た(1.2721g、4.071mmol、58%、無色の油状物)。出発物質94.3mg(0.221mmol、3%)、ジオール520.7mg(2.627mmol、37%)がそれぞれ回収された。
HF · pyridine complex (70%, 1.2 ml) was dissolved in pyridine (5.5 ml) and the solution was dissolved in (2R) -1,2-bis (tert-butyldimethylsilyloxy)-in THF (20 ml). A solution of 3- (4-methoxyphenoxy) propane (2.9927 g, 7.013 mmol) was added and stirred at room temperature for 13 hours. The reaction mixture was diluted with AcOEt (20 ml), washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane: AcOEt = 4: 1) to give the title compound (1.2721 g, 4.071 mmol, 58%, colorless oil). 94.3 mg (0.221 mmol, 3%) starting material and 520.7 mg (2.627 mmol, 37%) diol were recovered respectively.
1H-NMR(CDCl3):δ=6.826 (4H, s), 4.086 (1H, m), 3.911 (2H, m), 3.767 (3H, s), 3.722 (2H, m), 1.970 (1H, brs), 0.913 (9H, s), 0.143 (3H, s), 0.124 (3H, s)。
HRMS (ESI, [M+Na]+):計算値:C16H28NaO4Si+: 335.1649;実測値:335.1621。
工程5:(2S,3S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)プロポキシ)-tert-ブトキシホスホリルオキシ]酪酸tert-ブチルエステルの調製 1 H-NMR (CDCl 3 ): δ = 6.826 (4H, s), 4.086 (1H, m), 3.911 (2H, m), 3.767 (3H, s), 3.722 (2H, m), 1.970 (1H, brs), 0.913 (9H, s), 0.143 (3H, s), 0.124 (3H, s).
HRMS (ESI, [M + Na] + ): Calculated: C 16 H 28 NaO 4 Si + : 335.1649; Found: 335.1621.
Step 5: (2S, 3S) -2-tert-butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) propoxy) -tert-butoxy [Phosphoryloxy] butyric acid tert-butyl ester
HRMS (ESI, [M+Na]+):計算値:C16H28NaO4Si+: 335.1649;実測値:335.1621。
工程5:(2S,3S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)プロポキシ)-tert-ブトキシホスホリルオキシ]酪酸tert-ブチルエステルの調製 1 H-NMR (CDCl 3 ): δ = 6.826 (4H, s), 4.086 (1H, m), 3.911 (2H, m), 3.767 (3H, s), 3.722 (2H, m), 1.970 (1H, brs), 0.913 (9H, s), 0.143 (3H, s), 0.124 (3H, s).
HRMS (ESI, [M + Na] + ): Calculated: C 16 H 28 NaO 4 Si + : 335.1649; Found: 335.1621.
Step 5: (2S, 3S) -2-tert-butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) propoxy) -tert-butoxy [Phosphoryloxy] butyric acid tert-butyl ester
(2S,3S)-3-[(ジイソプロピルアミノ)-tert-ブトキシホスフィノオキシ]-2-(tert-ブトキシカルボニルアミノ)-3-メチル-プロピオン酸tert-ブチルエステル(298.2mg、0.623mmol)をCH2Cl2(5ml)およびトルエン(0.5ml)に溶解させ、減圧下溶媒を留去した。残渣に(2R)-2-(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)-1-プロパノール(232.1mg、0.743mmol)、CH2Cl2(5ml)およびトルエン(0.5ml)を加え、減圧下溶媒を留去した。アルゴン雰囲気下で残渣をCH2Cl2(5ml)に溶解させ、THF(5ml)中の1H-テトラゾール(136.9mg、1.954mmol)の溶液を室温で加えたところ、数分後に白色の固体が析出した。反応混合物を室温で3時間攪拌した後、tert-ブチルヒドロペルオキシド(TBHP)デカン溶液(5.0-6.0M、0.251ml、1.255mmol)を室温で加え、室温で1.5時間攪拌した。反応混合物を水(10ml)で希釈し、CH2Cl2(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=5:1)により精製し、表題の化合物を得た(341.9mg、0.484mmol、78%、無色の油状物)。
(2S, 3S) -3-[(Diisopropylamino) -tert-butoxyphosphinooxy] -2- (tert-butoxycarbonylamino) -3-methyl-propionic acid tert-butyl ester (298.2 mg, 0.623 mmol) ) Was dissolved in CH 2 Cl 2 (5 ml) and toluene (0.5 ml), and the solvent was distilled off under reduced pressure. To the residue was added (2R) -2- (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) -1-propanol (232.1 mg, 0.743 mmol), CH 2 Cl 2 (5 ml) and toluene (0 0.5 ml) and the solvent was distilled off under reduced pressure. The residue was dissolved in CH 2 Cl 2 (5 ml) under an argon atmosphere and a solution of 1H-tetrazole (136.9 mg, 1.954 mmol) in THF (5 ml) was added at room temperature, after a few minutes a white solid Precipitated. After the reaction mixture was stirred at room temperature for 3 hours, tert-butyl hydroperoxide (TBHP) decane solution (5.0-6.0 M, 0.251 ml, 1.255 mmol) was added at room temperature, and the mixture was stirred at room temperature for 1.5 hours. did. The reaction mixture was diluted with water (10 ml) and extracted with CH 2 Cl 2 (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane: AcOEt = 5: 1) to give the title compound (341.9 mg, 0.484 mmol, 78%, colorless oil).
1H-NMR(CDCl3):δ=6.825-6.816 (4H, m), 5.640 (1H, m), 4.646 (1H, m), 4.281 (1H, m), 4.180 (1H, m), 4.138 (1H, m), 4.006 (2H, m), 3.864 (1H, m), 3.764 (3H, s), 1.500-1.487 (9H, m), 1.468 (9H, s), 1.431 (12H, m), 0.893 (9H, s), 0.132 (3H, s), 0.108 (3H, s)。
31P-NMR(CDCl3):δ=-6.220, -5.903。 1 H-NMR (CDCl 3 ): δ = 6.825-6.816 (4H, m), 5.640 (1H, m), 4.646 (1H, m), 4.281 (1H, m), 4.180 (1H, m), 4.138 ( 1H, m), 4.006 (2H, m), 3.864 (1H, m), 3.764 (3H, s), 1.500-1.487 (9H, m), 1.468 (9H, s), 1.431 (12H, m), 0.893 (9H, s), 0.132 (3H, s), 0.108 (3H, s).
31 P-NMR (CDCl 3 ): δ = -6.220, -5.903.
31P-NMR(CDCl3):δ=-6.220, -5.903。 1 H-NMR (CDCl 3 ): δ = 6.825-6.816 (4H, m), 5.640 (1H, m), 4.646 (1H, m), 4.281 (1H, m), 4.180 (1H, m), 4.138 ( 1H, m), 4.006 (2H, m), 3.864 (1H, m), 3.764 (3H, s), 1.500-1.487 (9H, m), 1.468 (9H, s), 1.431 (12H, m), 0.893 (9H, s), 0.132 (3H, s), 0.108 (3H, s).
31 P-NMR (CDCl 3 ): δ = -6.220, -5.903.
工程6:(2S,3S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-ヒドロキシプロポキシ)-tert-ブトキシホスホリルオキシ]酪酸tert-ブチルエステルの調製
Step 6: (2S, 3S) -2-tert-butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3-hydroxypropoxy) -tert-butoxyphosphoryloxy] butyric acid tert -Preparation of butyl esters
(2S,3S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)プロポキシ)-tert-ブトキシホスホリルオキシ]酪酸(300.3mg、0.425mmol)をCH3CN(8ml)およびH2O(2ml)に溶解させ、0℃で撹拌し、硝酸セリウムアンモニウム(581.8mg、1.061mmol)を加え、0℃で1時間攪拌した。H2O(10ml)を加えて反応をクエンチし、有機層を食塩水で洗浄し、Na2SO4で乾燥し、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=1:1)により精製し、表題の化合物を得た(177.4mg、0.296mmol、70%、褐色の油状物)。
(2S, 3S) -2-tert-butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) propoxy) -tert-butoxyphosphoryloxy] Butyric acid (300.3 mg, 0.425 mmol) was dissolved in CH 3 CN (8 ml) and H 2 O (2 ml), stirred at 0 ° C., ceric ammonium nitrate (581.8 mg, 1.061 mmol) was added, and 0 Stir at 1 ° C. for 1 hour. H 2 O (10 ml) was added to quench the reaction, the organic layer was washed with brine, dried over Na 2 SO 4 and the solvent was evaporated. The residue was purified by column chromatography (hexane: AcOEt = 1: 1) to give the title compound (177.4 mg, 0.296 mmol, 70%, brown oil).
1H-NMR(CDCl3):δ=5.621 (1H, m), 4.683 (1H, m), 4.305 (1H, m), 4.026 (1H, m), 3.973 (1H, m), 3.911 (1H, m), 3.630 (2H, m), 1.488 (18H, s), 1.445 (12H, m), 0.986 (9H, s), 0.146 (6H, s)。
1 H-NMR (CDCl 3 ): δ = 5.621 (1H, m), 4.683 (1H, m), 4.305 (1H, m), 4.026 (1H, m), 3.973 (1H, m), 3.911 (1H, m), 3.630 (2H, m), 1.488 (18H, s), 1.445 (12H, m), 0.986 (9H, s), 0.146 (6H, s).
工程7:(2S,3S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-ヒドロキシプロポキシ)-tert-ブトキシホスホリルオキシ]酪酸tert-ブチルエステルの調製
Step 7: (2S, 3S) -2-tert-Butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3-hydroxypropoxy) -tert-butoxyphosphoryloxy] butyric acid tert -Preparation of butyl esters
(2S,3S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-ヒドロキシプロポキシ)-tert-ブトキシホスホリルオキシ]酪酸(98.7mg、0.165mmol)をCH2Cl2(2ml)に溶解させ、4-ジメチルアミノピリジン(60.3mg、0.502mmol)を加え、0℃に冷却した。当該混合物に、CH2Cl2(1ml)中のオレオイルクロリド(69.9mg、0.232mmol)の溶液を0℃で滴下して加え、13時間攪拌した。反応混合物にMeOH(1ml)を加え、2時間攪拌し、濃縮した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=3:1)により精製し、表題の化合物を得た(122.2mg、0.141mmol、86%、黄色油状物)。
(2S, 3S) -2-tert-Butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3-hydroxypropoxy) -tert-butoxyphosphoryloxy] butyric acid (98.7 mg , 0.165 mmol) was dissolved in CH 2 Cl 2 (2 ml), 4-dimethylaminopyridine (60.3 mg, 0.502 mmol) was added and cooled to 0 ° C. To the mixture was added dropwise a solution of oleoyl chloride (69.9 mg, 0.232 mmol) in CH 2 Cl 2 (1 ml) at 0 ° C. and stirred for 13 hours. To the reaction mixture was added MeOH (1 ml), stirred for 2 hours and concentrated. The residue was purified by column chromatography (hexane: AcOEt = 3: 1) to give the title compound (122.2 mg, 0.141 mmol, 86%, yellow oil).
1H-NMR(CDCl3):δ=5.642 (1H, m), 5.336 (2H, m), 4.660 (1H, m), 4.277 (1H, m), 4.155 (1H, m), 4.023 (2H, m), 3.908 (2H, m), 2.304 (2H, t, J=7.44 Hz), 1.999 (4H, m), 1.608 (2H, m), 1.476 (18H, m), 1.433 (12H, m), 1.272 (20H, m), 0.873 (12H, m), 0.100 (3H, s), 0.089 (3H, s)。
13C-NMR(CDCl3):δ=173.46, 167.88, 155.37, 129.98, 129.72, 82.50, 79.79, 75.71, 69.15, 69.06, 67.79, 67.73, 65.03, 64.99, 58.62, 34.14, 31.88, 29.82, 29.78, 29.74, 29.70, 29.50, 29.30, 29.18, 29.13, 29.10, 28.91, 28.31, 28.03, 27.20, 27.16, 25.66, 24.86, 22.66, 18.75, 18.59, 18.02, 14.10, -4.74。
31P-NMR(CDCl3):δ=-5.961, -6.161。
HRMS (ESI, [M+Na]+):計算値:C44H86NNaO11PSi+: 886.5606;実測値:886.5592。 1 H-NMR (CDCl 3 ): δ = 5.642 (1H, m), 5.336 (2H, m), 4.660 (1H, m), 4.277 (1H, m), 4.155 (1H, m), 4.023 (2H, m), 3.908 (2H, m), 2.304 (2H, t, J = 7.44 Hz), 1.999 (4H, m), 1.608 (2H, m), 1.476 (18H, m), 1.433 (12H, m), 1.272 (20H, m), 0.873 (12H, m), 0.100 (3H, s), 0.089 (3H, s).
13 C-NMR (CDCl 3 ): δ = 173.46, 167.88, 155.37, 129.98, 129.72, 82.50, 79.79, 75.71, 69.15, 69.06, 67.79, 67.73, 65.03, 64.99, 58.62, 34.14, 31.88, 29.82, 29.78, 29.74 , 29.70, 29.50, 29.30, 29.18, 29.13, 29.10, 28.91, 28.31, 28.03, 27.20, 27.16, 25.66, 24.86, 22.66, 18.75, 18.59, 18.02, 14.10, -4.74.
31 P-NMR (CDCl 3 ): δ = −5.961, −6.161.
HRMS (ESI, [M + Na] + ): Calculated: C 44 H 86 NNaO 11 PSi + : 886.5606; Found: 886.5592.
13C-NMR(CDCl3):δ=173.46, 167.88, 155.37, 129.98, 129.72, 82.50, 79.79, 75.71, 69.15, 69.06, 67.79, 67.73, 65.03, 64.99, 58.62, 34.14, 31.88, 29.82, 29.78, 29.74, 29.70, 29.50, 29.30, 29.18, 29.13, 29.10, 28.91, 28.31, 28.03, 27.20, 27.16, 25.66, 24.86, 22.66, 18.75, 18.59, 18.02, 14.10, -4.74。
31P-NMR(CDCl3):δ=-5.961, -6.161。
HRMS (ESI, [M+Na]+):計算値:C44H86NNaO11PSi+: 886.5606;実測値:886.5592。 1 H-NMR (CDCl 3 ): δ = 5.642 (1H, m), 5.336 (2H, m), 4.660 (1H, m), 4.277 (1H, m), 4.155 (1H, m), 4.023 (2H, m), 3.908 (2H, m), 2.304 (2H, t, J = 7.44 Hz), 1.999 (4H, m), 1.608 (2H, m), 1.476 (18H, m), 1.433 (12H, m), 1.272 (20H, m), 0.873 (12H, m), 0.100 (3H, s), 0.089 (3H, s).
13 C-NMR (CDCl 3 ): δ = 173.46, 167.88, 155.37, 129.98, 129.72, 82.50, 79.79, 75.71, 69.15, 69.06, 67.79, 67.73, 65.03, 64.99, 58.62, 34.14, 31.88, 29.82, 29.78, 29.74 , 29.70, 29.50, 29.30, 29.18, 29.13, 29.10, 28.91, 28.31, 28.03, 27.20, 27.16, 25.66, 24.86, 22.66, 18.75, 18.59, 18.02, 14.10, -4.74.
31 P-NMR (CDCl 3 ): δ = −5.961, −6.161.
HRMS (ESI, [M + Na] + ): Calculated: C 44 H 86 NNaO 11 PSi + : 886.5606; Found: 886.5592.
工程8:(2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸の調製
Step 8: Preparation of (2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid
(2S,3S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-ヒドロキシプロポキシ)-tert-ブトキシホスホリルオキシ]酪酸tert-ブチルエステル(87.4mg、0.101mmol)を0℃でトリフルオロ酢酸(1.0ml)に溶解させ、室温で1時間攪拌した。反応混合物を濃縮し、残渣をカラムクロマトグラフィー(CHCl3/MeOH/AcOH=8:1:1→6:1:3)により精製し、表題の化合物を酢酸塩として得た(43.7mg、0.0813mmol、80%、白色粉末)。得られた酢酸塩をトリフルオロ酢酸に溶解させ、溶媒を除去して表題の化合物のトリフルオロ酢酸塩を得た。
(2S, 3S) -2-tert-Butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3-hydroxypropoxy) -tert-butoxyphosphoryloxy] butyric acid tert-butyl ester (87.4 mg, 0.101 mmol) was dissolved in trifluoroacetic acid (1.0 ml) at 0 ° C. and stirred at room temperature for 1 hour. The reaction mixture was concentrated and the residue was purified by column chromatography (CHCl 3 / MeOH / AcOH = 8: 1: 1 → 6: 1: 3) to give the title compound as an acetate salt (43.7 mg, 0 .0813 mmol, 80%, white powder). The resulting acetate was dissolved in trifluoroacetic acid and the solvent was removed to give the trifluoroacetate of the title compound.
1H-NMR(CDCl3/TFA-d=4:1):δ=5.362 (2H, m), 5.135 (1H, m), 4.449-4.179 (6H, m), 2.445 (2H, m), 2.017 (4H, m), 1.624 (5H,m), 1.284 (20H, m), 0.874 (3H, m)。
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.816。
HRMS (MALDI-TOF, [M+Na]+):Calcd for C25H48NNaO9P+: 560.2959. Found: 560.2912。
mp. 197.0℃-198.5℃。
元素分析:計算値:C25H48NO9P・1.8CF3COOH: C, 46.24; H, 6.76; N, 1.89;実測値:C, 46.42; H, 6.85; N, 1.88。
上記実施例と同様の手法により、以下の化合物を調製した。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.362 (2H, m), 5.135 (1H, m), 4.449-4.179 (6H, m), 2.445 (2H, m), 2.017 (4H, m), 1.624 (5H, m), 1.284 (20H, m), 0.874 (3H, m).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.816.
HRMS (MALDI-TOF, [M + Na] + ): Calcd for C 25 H 48 NNaO 9 P + : 560.2959. Found: 560.2912.
mp. 197.0 ℃ -198.5 ℃.
Calcd: C 25 H 48 NO 9 P · 1.8CF 3 COOH: C, 46.24; H, 6.76; N, 1.89; Found: C, 46.42; H, 6.85 ; N, 1.88.
The following compounds were prepared in the same manner as in the above examples.
31P-NMR(CDCl3/TFA-d=4:1):δ=-1.816。
HRMS (MALDI-TOF, [M+Na]+):Calcd for C25H48NNaO9P+: 560.2959. Found: 560.2912。
mp. 197.0℃-198.5℃。
元素分析:計算値:C25H48NO9P・1.8CF3COOH: C, 46.24; H, 6.76; N, 1.89;実測値:C, 46.42; H, 6.85; N, 1.88。
上記実施例と同様の手法により、以下の化合物を調製した。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 5.362 (2H, m), 5.135 (1H, m), 4.449-4.179 (6H, m), 2.445 (2H, m), 2.017 (4H, m), 1.624 (5H, m), 1.284 (20H, m), 0.874 (3H, m).
31 P-NMR (CDCl 3 / TFA-d = 4: 1): δ = -1.816.
HRMS (MALDI-TOF, [M + Na] + ): Calcd for C 25 H 48 NNaO 9 P + : 560.2959. Found: 560.2912.
mp. 197.0 ℃ -198.5 ℃.
Calcd: C 25 H 48 NO 9 P · 1.8
The following compounds were prepared in the same manner as in the above examples.
[実施例25](2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸
[Example 25] (2S, 3S) -2-Amino-3-[((2R) -2-hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid
1H-NMR(CDCl3):δ=5.086 (1H, m), 4.433 (1H, m), 4.376 (1H, m), 4.282 (3H, m), 4.134 (1H, m), 2.430 (2H, t, J=7.52 Hz), 1.619 (5H, m), 1.260 (25H, m), 0.875 (3H, t, J=6.60 Hz)。
31P-NMR(CDCl3):δ=-1.649。
HRMS (MALDI-TOF, [M+Na]+): Calcd for C23H46NNaO9P+: 534.3802. Found: 534.2806。
融点:193.2℃-194.5℃。
元素分析:計算値:C23H46NO9P・5CF3COOH・4H2O: C, 34.35; H, 5.15; N, 1.21;実測値:C, 34.14; H, 5.28; N, 1.49。 1 H-NMR (CDCl 3 ): δ = 5.086 (1H, m), 4.433 (1H, m), 4.376 (1H, m), 4.282 (3H, m), 4.134 (1H, m), 2.430 (2H, t, J = 7.52 Hz), 1.619 (5H, m), 1.260 (25H, m), 0.875 (3H, t, J = 6.60 Hz).
31 P-NMR (CDCl 3 ): δ = -1.649.
HRMS (MALDI-TOF, [M + Na] + ): Calcd for C 23 H 46 NNaO 9 P + : 534.3802. Found: 534.2806.
Melting point: 193.2 ° C-194.5 ° C.
Calcd: C 23 H 46 NO 9 P ·5CF 3 COOH · 4H 2 O: C, 34.35; H, 5.15; N, 1.21; Found: C, 34.14; H, 5.28 ; N, 1.49.
31P-NMR(CDCl3):δ=-1.649。
HRMS (MALDI-TOF, [M+Na]+): Calcd for C23H46NNaO9P+: 534.3802. Found: 534.2806。
融点:193.2℃-194.5℃。
元素分析:計算値:C23H46NO9P・5CF3COOH・4H2O: C, 34.35; H, 5.15; N, 1.21;実測値:C, 34.14; H, 5.28; N, 1.49。 1 H-NMR (CDCl 3 ): δ = 5.086 (1H, m), 4.433 (1H, m), 4.376 (1H, m), 4.282 (3H, m), 4.134 (1H, m), 2.430 (2H, t, J = 7.52 Hz), 1.619 (5H, m), 1.260 (25H, m), 0.875 (3H, t, J = 6.60 Hz).
31 P-NMR (CDCl 3 ): δ = -1.649.
HRMS (MALDI-TOF, [M + Na] + ): Calcd for C 23 H 46 NNaO 9 P + : 534.3802. Found: 534.2806.
Melting point: 193.2 ° C-194.5 ° C.
Calcd: C 23 H 46 NO 9 P ·
以下の化合物はJ. Med. Chem. 2009, 52, 5837-5863.に記載の方法により調製した。
The following compounds were prepared by the method described in J. Med. Chem. 2009, 52, 5837-5863.
[実施例26](2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸
[Example 26] (2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid
1H-NMR(CDCl3):δ=4.23 (2H, m), 4.05 (5H, m), 2.24 (2H, m), 1.52 (2H, m), 1.17 (31H, m), 0.79 (3H, t, J=6.8 Hz)。
HRMS (ESI, [M-H]-): Calcd for C25H49NO9P-: 538.3145. Found: 538.3137。
融点:179.0℃-179.5℃。 1 H-NMR (CDCl 3 ): δ = 4.23 (2H, m), 4.05 (5H, m), 2.24 (2H, m), 1.52 (2H, m), 1.17 (31H, m), 0.79 (3H, t, J = 6.8 Hz).
HRMS (ESI, [MH] - ): Calcd for C 25 H 49 NO 9 P -: 538.3145. Found: 538.3137.
Melting point: 179.0 ° C-179.5 ° C.
HRMS (ESI, [M-H]-): Calcd for C25H49NO9P-: 538.3145. Found: 538.3137。
融点:179.0℃-179.5℃。 1 H-NMR (CDCl 3 ): δ = 4.23 (2H, m), 4.05 (5H, m), 2.24 (2H, m), 1.52 (2H, m), 1.17 (31H, m), 0.79 (3H, t, J = 6.8 Hz).
HRMS (ESI, [MH] - ): Calcd for C 25 H 49 NO 9 P -: 538.3145. Found: 538.3137.
Melting point: 179.0 ° C-179.5 ° C.
出発原料として(S)-(-)-2,2-ジメチル-1,3-ジオキソラン-4-メタノールを用い、中間体として(2S)-3-[(ジイソプロピルアミノ)-tert-ブトキシホスフィノオキシ]-2-(tert-ブトキシカルボニルアミノ)-3-メチル-プロピオン酸tert-ブチルエステルを用いて、実施例24と同様の手法で以下の化合物を調製した。
(S)-(−)-2,2-dimethyl-1,3-dioxolane-4-methanol was used as a starting material, and (2S) -3-[(diisopropylamino) -tert-butoxyphosphinooxy was used as an intermediate. ] The following compound was prepared in the same manner as in Example 24 using 2- (tert-butoxycarbonylamino) -3-methyl-propionic acid tert-butyl ester.
[実施例27](2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 27] (2S) -2-Amino-3-[((2S) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3):δ= 5.367 (2H, m), 4.671 (2H, m), 4.546 (1H, m), 4.281 (5H, m), 2.423 (2H, m), 2.018 (4H, m), 1.609 (2H, m), 1.270 (20H, m), 0.872 (3H, t, J=6.4 Hz)。
31P-NMR(CDCl3):δ=-1.147。
HRMS (MALDI-TOF, [M+Na]+):計算値:C24H46NNaO9P+:546.2802;実測値:546.2784。
融点:131.0-131.9℃。
以下の化合物はJ. Med. Chem. 2009, 52, 5837-5863.に記載の方法により調製した。 1 H-NMR (CDCl 3 ): δ = 5.367 (2H, m), 4.671 (2H, m), 4.546 (1H, m), 4.281 (5H, m), 2.423 (2H, m), 2.018 (4H, m), 1.609 (2H, m), 1.270 (20H, m), 0.872 (3H, t, J = 6.4 Hz).
31 P-NMR (CDCl 3 ): δ = -1.147.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 24 H 46 NNaO 9 P + : 546.2802; Found: 546.2784.
Melting point: 131.0-131.9 ° C.
The following compounds were prepared by the method described in J. Med. Chem. 2009, 52, 5837-5863.
31P-NMR(CDCl3):δ=-1.147。
HRMS (MALDI-TOF, [M+Na]+):計算値:C24H46NNaO9P+:546.2802;実測値:546.2784。
融点:131.0-131.9℃。
以下の化合物はJ. Med. Chem. 2009, 52, 5837-5863.に記載の方法により調製した。 1 H-NMR (CDCl 3 ): δ = 5.367 (2H, m), 4.671 (2H, m), 4.546 (1H, m), 4.281 (5H, m), 2.423 (2H, m), 2.018 (4H, m), 1.609 (2H, m), 1.270 (20H, m), 0.872 (3H, t, J = 6.4 Hz).
31 P-NMR (CDCl 3 ): δ = -1.147.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 24 H 46 NNaO 9 P + : 546.2802; Found: 546.2784.
Melting point: 131.0-131.9 ° C.
The following compounds were prepared by the method described in J. Med. Chem. 2009, 52, 5837-5863.
[実施例28](2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 28] (2S) -2-Amino-3-[((2S) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3):δ=4.59 (1H, m), 4.34 (1H, m), 4.17 (1H, m), 4.04 (2H, m), 3.90 (2H, m), 3.72 (1H, m), 2.23 (2H, m), 1.55 (2H, m), 1.08 (28H, m), 0.79 (3H, t, J=7.0 Hz)。
HRMS (ESI, [M-H]-): Calcd for C24H47NO9P-: 524.2988. Found: 524.2971。
融点:171.5℃-172.0℃。
元素分析:計算値:C24H47NO9P・0.8CF3COOH: C, 50.57; H, 8.15; N, 2.33;実測値:C, 50.31; H, 7.82; N, 2.32。 1 H-NMR (CDCl 3 ): δ = 4.59 (1H, m), 4.34 (1H, m), 4.17 (1H, m), 4.04 (2H, m), 3.90 (2H, m), 3.72 (1H, m), 2.23 (2H, m), 1.55 (2H, m), 1.08 (28H, m), 0.79 (3H, t, J = 7.0 Hz).
HRMS (ESI, [MH] - ): Calcd for C 24 H 47 NO 9 P -:. 524.2988 Found: 524.2971.
Melting point: 171.5 ° C-172.0 ° C.
Calcd: C 24 H 47 NO 9 P · 0.8CF 3 COOH: C, 50.57; H, 8.15; N, 2.33; Found: C, 50.31; H, 7.82 ; N, 2.32.
HRMS (ESI, [M-H]-): Calcd for C24H47NO9P-: 524.2988. Found: 524.2971。
融点:171.5℃-172.0℃。
元素分析:計算値:C24H47NO9P・0.8CF3COOH: C, 50.57; H, 8.15; N, 2.33;実測値:C, 50.31; H, 7.82; N, 2.32。 1 H-NMR (CDCl 3 ): δ = 4.59 (1H, m), 4.34 (1H, m), 4.17 (1H, m), 4.04 (2H, m), 3.90 (2H, m), 3.72 (1H, m), 2.23 (2H, m), 1.55 (2H, m), 1.08 (28H, m), 0.79 (3H, t, J = 7.0 Hz).
HRMS (ESI, [MH] - ): Calcd for C 24 H 47 NO 9 P -:. 524.2988 Found: 524.2971.
Melting point: 171.5 ° C-172.0 ° C.
Calcd: C 24 H 47 NO 9 P · 0.8
[実施例29](2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 29] (2S) -2-Amino-3-[((2S) -2-hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3):δ=4.684 (2H, m), 4.544 (1H, m), 4.282 (5H, m), 2.426 (2H, t, J=7.8 Hz), 1.609 (2H, m), 1.257 (24H, m), 0.874 (3H, t, J=6.8 Hz)。
31P-NMR(CDCl3):δ=-1.272。
HRMS (MALDI-TOF, [M+Na]+):計算値:C22H44NNaO9P+:520.2646;実測値:520.2644。
融点:128.1-129.2℃。 1 H-NMR (CDCl 3 ): δ = 4.684 (2H, m), 4.544 (1H, m), 4.282 (5H, m), 2.426 (2H, t, J = 7.8 Hz), 1.609 (2H, m) , 1.257 (24H, m), 0.874 (3H, t, J = 6.8 Hz).
31 P-NMR (CDCl 3 ): δ = -1.272.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 22 H 44 NNaO 9 P + : 520.2646; Found: 520.2644.
Melting point: 128.1-129.2 ° C.
31P-NMR(CDCl3):δ=-1.272。
HRMS (MALDI-TOF, [M+Na]+):計算値:C22H44NNaO9P+:520.2646;実測値:520.2644。
融点:128.1-129.2℃。 1 H-NMR (CDCl 3 ): δ = 4.684 (2H, m), 4.544 (1H, m), 4.282 (5H, m), 2.426 (2H, t, J = 7.8 Hz), 1.609 (2H, m) , 1.257 (24H, m), 0.874 (3H, t, J = 6.8 Hz).
31 P-NMR (CDCl 3 ): δ = -1.272.
HRMS (MALDI-TOF, [M + Na] + ): Calculated: C 22 H 44 NNaO 9 P + : 520.2646; Found: 520.2644.
Melting point: 128.1-129.2 ° C.
[実施例30](2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(2-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 30] (2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (2-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid
工程1:3-(2-ヒドロキシフェニル)プロピオン酸メチルエステル
Step 1: 3- (2-hydroxyphenyl) propionic acid methyl ester
2-(ヒドロキシフェニル)プロピオン酸(1.0030g、6.036mmol)およびKHCO3(963.8mg、9.626mmol)をDMF(10ml)に加え、さらにヨウ化メチル(1.3672g、9.632mmol)を加え、室温で5時間攪拌した。反応混合物に水(10ml)を加え、AcOEt(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=2:1)により精製し、表題の化合物を得た(1.0243g、5.684mmol、94%、白色固体)。
2- (Hydroxyphenyl) propionic acid (1.0030 g, 6.036 mmol) and KHCO 3 (963.8 mg, 9.626 mmol) were added to DMF (10 ml), followed by methyl iodide (1.3672 g, 9.632 mmol). And stirred at room temperature for 5 hours. Water (10 ml) was added to the reaction mixture, and the mixture was extracted with AcOEt (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 2: 1) to give the title compound (1.0243 g, 5.684 mmol, 94%, white solid).
1H-NMR(CDCl3):δ=7.143-7.077 (2H, m), 6.997 (1H, s), 6.879 (1H, d, J=7.68 Hz), 6.869 (1H, t, J=7.36 Hz), 3.691 (3H, s), 2.909 (2H, t, J=6.42 Hz), 2.725 (2H, t, J=6.42 Hz)。
1 H-NMR (CDCl 3 ): δ = 7.143-7.077 (2H, m), 6.997 (1H, s), 6.879 (1H, d, J = 7.68 Hz), 6.869 (1H, t, J = 7.36 Hz) , 3.691 (3H, s), 2.909 (2H, t, J = 6.42 Hz), 2.725 (2H, t, J = 6.42 Hz).
工程2:3-(2-ヘプチルオキシフェニル)プロピオン酸メチルエステル
Step 2: 3- (2-Heptyloxyphenyl) propionic acid methyl ester
NaH(84.5mg、2.113mmol)をDMF(2ml)中に懸濁させ0℃で撹拌し、DMF(2ml)中の3-(2-ヒドロキシフェニル)プロピオン酸メチルエステルの溶液(256.0mg、1.421mmol)を滴下して加え、30分間攪拌した。反応混合物にDMF(2ml)中の1-ブロモヘプタン(372.8mg、2.082mmol)の溶液を滴下して加え、室温で25時間攪拌した。反応混合物をH2O(10ml)で希釈し、AcOEt(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=10:0→9:1)により精製し、表題の化合物を得た(232.8mg、0.836mmol、59%、無色の油状物)。
NaH (84.5 mg, 2.113 mmol) was suspended in DMF (2 ml) and stirred at 0 ° C., and a solution of 3- (2-hydroxyphenyl) propionic acid methyl ester in DMF (2 ml) (256.0 mg). , 1.421 mmol) was added dropwise and stirred for 30 minutes. To the reaction mixture was added dropwise a solution of 1-bromoheptane (372.8 mg, 2.082 mmol) in DMF (2 ml) and stirred at room temperature for 25 hours. The reaction mixture was diluted with H 2 O (10 ml) and extracted with AcOEt (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 10: 0 → 9: 1) to give the title compound (232.8 mg, 0.836 mmol, 59%, colorless oil).
1H NMR (CDCl3) :δ=7.192-7.136 (2H, m), 6.875-6.814 (2H, m), 3.960 (2H, t, J=6.44 Hz), 3.668 (3H, s), 2.946 (2H, t, J=7.80 Hz), 2.625 (2H, t, J=7.80 Hz), 1.799 (2H, m), 1.453 (2H, m), 1.400-1.316 (6H, m), 0.900 (3H, t, J=6.50 Hz)。
1 H NMR (CDCl 3 ): δ = 7.192-7.136 (2H, m), 6.875-6.814 (2H, m), 3.960 (2H, t, J = 6.44 Hz), 3.668 (3H, s), 2.946 (2H , t, J = 7.80 Hz), 2.625 (2H, t, J = 7.80 Hz), 1.799 (2H, m), 1.453 (2H, m), 1.400-1.316 (6H, m), 0.900 (3H, t, J = 6.50 Hz).
工程3:3-(2-ヘプチルオキシフェニル)プロピオン酸
Step 3: 3- (2-Heptyloxyphenyl) propionic acid
3-(2-ヘプチルオキシフェニル)プロピオン酸メチルエステル(206.0mg、0.740mmol)をMeOH(5ml)およびTHF(5ml)に溶解させ、NaOH水溶液(2N、5ml)を加え、室温で2時間攪拌した。HCl水溶液(1N)を加えて水層のpHを2にしてAcOEt(20ml)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン/AcOEt=1:1)により精製し、表題の化合物を得た(187.8mg、0.710mmol、96%、白色固体)。
3- (2-Heptyloxyphenyl) propionic acid methyl ester (206.0 mg, 0.740 mmol) was dissolved in MeOH (5 ml) and THF (5 ml), aqueous NaOH (2N, 5 ml) was added, and the mixture was stirred at room temperature for 2 hours. Stir. Aqueous HCl (1N) was added to adjust the pH of the aqueous layer to 2, and the mixture was extracted with AcOEt (20 ml). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane / AcOEt = 1: 1) to give the title compound (187.8 mg, 0.710 mmol, 96%, white solid).
1H NMR (CDCl3) :δ=11.361 (1H, brs), 7.206-7.153 (2H, m), 6.883-6.822 (2H, m), 3.967 (2H, t, J=6.46 Hz), 2.957 (2H, t, J=7.76 Hz), 2.679 (2H, t, J=7.78 Hz), 1.806 (2H, m), 1.496 (2H, m), 1.388-1.313 (6H, m), 0.902 (3H, t, J=6.80 Hz)。
13C NMR (CDCl3) :δ=179.28, 156.96, 129.94, 128.57, 127.63, 120.19, 110.98, 67.75, 33.90, 31.77, 29.31, 29.03, 26.11, 25.98, 22.59, 14.07。
HRMS (ESI, [M-H]-):Calcd for C16H23O3 -: 263.1653. Found: 263.1552。
融点:< 30 ℃。 1 H NMR (CDCl 3 ): δ = 11.361 (1H, brs), 7.206-7.153 (2H, m), 6.883-6.822 (2H, m), 3.967 (2H, t, J = 6.46 Hz), 2.957 (2H , t, J = 7.76 Hz), 2.679 (2H, t, J = 7.78 Hz), 1.806 (2H, m), 1.496 (2H, m), 1.388-1.313 (6H, m), 0.902 (3H, t, J = 6.80 Hz).
13 C NMR (CDCl 3 ): δ = 179.28, 156.96, 129.94, 128.57, 127.63, 120.19, 110.98, 67.75, 33.90, 31.77, 29.31, 29.03, 26.11, 25.98, 22.59, 14.07.
HRMS (ESI, [MH] − ): Calcd for C 16 H 23 O 3 − : 263.1653. Found: 263.1552.
Melting point: <30 ° C.
13C NMR (CDCl3) :δ=179.28, 156.96, 129.94, 128.57, 127.63, 120.19, 110.98, 67.75, 33.90, 31.77, 29.31, 29.03, 26.11, 25.98, 22.59, 14.07。
HRMS (ESI, [M-H]-):Calcd for C16H23O3 -: 263.1653. Found: 263.1552。
融点:< 30 ℃。 1 H NMR (CDCl 3 ): δ = 11.361 (1H, brs), 7.206-7.153 (2H, m), 6.883-6.822 (2H, m), 3.967 (2H, t, J = 6.46 Hz), 2.957 (2H , t, J = 7.76 Hz), 2.679 (2H, t, J = 7.78 Hz), 1.806 (2H, m), 1.496 (2H, m), 1.388-1.313 (6H, m), 0.902 (3H, t, J = 6.80 Hz).
13 C NMR (CDCl 3 ): δ = 179.28, 156.96, 129.94, 128.57, 127.63, 120.19, 110.98, 67.75, 33.90, 31.77, 29.31, 29.03, 26.11, 25.98, 22.59, 14.07.
HRMS (ESI, [MH] − ): Calcd for C 16 H 23 O 3 − : 263.1653. Found: 263.1552.
Melting point: <30 ° C.
工程4:(2S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)プロポキシ)-tert-ブトキシホスホリルオキシ]プロピオン酸tert-ブチルエステルの調製
Step 4: (2S) -2-tert-butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) propoxy) -tert-butoxyphosphoryloxy ] Preparation of propionic acid tert-butyl ester
(2S)-3-[(ジイソプロピルアミノ)-tert-ブトキシホスフィノオキシ]-2-(tert-ブトキシカルボニルアミノ)プロピオン酸tert-ブチルエステル(498.7mg、1.074mmol)をCH2Cl2(5ml)およびトルエン(0.5ml)の混合液に溶解させ、減圧下溶媒を留去した。残渣に、(2R)-2-(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)-1-プロパノール(423.5mg、1.355mmol)、CH2Cl2(5ml)およびトルエン(0.5ml)を加え、減圧下溶媒を留去した。アルゴン雰囲気下で残渣をCH2Cl2(7ml)に溶解させ、THF(7ml)中の1H-テトラゾール(226.0mg、3.226mmol)の溶液を室温で加えたところ、数分後に白色の固体が析出した。反応混合物を室温で3.5時間攪拌し、tert-ブチルヒドロペルオキシド(TBHP)デカン溶液(5.0-6.0M、0.431ml、2.155mmol)を室温で加え、さらに1.5時間攪拌した。反応混合物を水(15ml)で希釈し、CH2Cl2(20ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥し、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=5:1)により精製し、表題の化合物を得た(783.3mg、1.132mmol、84%、無色の油状物)。
(2S) -3-[(Diisopropylamino) -tert-butoxyphosphinooxy] -2- (tert-butoxycarbonylamino) propionic acid tert-butyl ester (498.7 mg, 1.074 mmol) was added to CH 2 Cl 2 ( 5 ml) and toluene (0.5 ml), and the solvent was distilled off under reduced pressure. To the residue was added (2R) -2- (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) -1-propanol (423.5 mg, 1.355 mmol), CH 2 Cl 2 (5 ml) and toluene ( 0.5 ml) and the solvent was distilled off under reduced pressure. The residue was dissolved in CH 2 Cl 2 (7 ml) under an argon atmosphere and a solution of 1H-tetrazole (226.0 mg, 3.226 mmol) in THF (7 ml) was added at room temperature, after a few minutes a white solid Precipitated. The reaction mixture was stirred at room temperature for 3.5 hours, tert-butyl hydroperoxide (TBHP) decane solution (5.0-6.0 M, 0.431 ml, 2.155 mmol) was added at room temperature, and the mixture was further stirred for 1.5 hours. did. The reaction mixture was diluted with water (15 ml) and extracted with CH 2 Cl 2 (20 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane: AcOEt = 5: 1) to give the title compound (783.3 mg, 1.132 mmol, 84%, colorless oil).
1H-NMR(CDCl3):δ=6.822 (4H, m), 5.482 (1H, m), 4.357 (1H, m), 4.256-4.148 (2H, m), 4.113-3.928 (3H, m), 3.854 (1H, m), 3.766 (3H, s), 1.484 (9H, s), 1.465 (9H, s), 1.435 (9H, s), 0.895 (9H, s), 0.129 (3H, s), 0.108 (3H, s)。
31P-NMR(CDCl3):δ=-5.584, -5.823。
HRMS (ESI, [M+Na]+):計算値:C32H58NNaO11PSi+: 714.3409;実測値:714.3389。 1 H-NMR (CDCl 3 ): δ = 6.822 (4H, m), 5.482 (1H, m), 4.357 (1H, m), 4.256-4.148 (2H, m), 4.113-3.928 (3H, m), 3.854 (1H, m), 3.766 (3H, s), 1.484 (9H, s), 1.465 (9H, s), 1.435 (9H, s), 0.895 (9H, s), 0.129 (3H, s), 0.108 (3H, s).
31 P-NMR (CDCl 3 ): δ = −5.584, −5.823.
HRMS (ESI, [M + Na] + ): Calculated: C 32 H 58 NNaO 11 PSi + : 714.3409; Found: 714.3389.
31P-NMR(CDCl3):δ=-5.584, -5.823。
HRMS (ESI, [M+Na]+):計算値:C32H58NNaO11PSi+: 714.3409;実測値:714.3389。 1 H-NMR (CDCl 3 ): δ = 6.822 (4H, m), 5.482 (1H, m), 4.357 (1H, m), 4.256-4.148 (2H, m), 4.113-3.928 (3H, m), 3.854 (1H, m), 3.766 (3H, s), 1.484 (9H, s), 1.465 (9H, s), 1.435 (9H, s), 0.895 (9H, s), 0.129 (3H, s), 0.108 (3H, s).
31 P-NMR (CDCl 3 ): δ = −5.584, −5.823.
HRMS (ESI, [M + Na] + ): Calculated: C 32 H 58 NNaO 11 PSi + : 714.3409; Found: 714.3389.
工程5:(2S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-ヒドロキシプロポキシ)-tert-ブトキシホスホリルオキシ]プロピオン酸tert-ブチルエステルの調製
Step 5: (2S) -2-tert-butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3-hydroxypropoxy) -tert-butoxyphosphoryloxy] propionic acid tert- Preparation of butyl ester
(2S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-(4-メトキシフェノキシ)プロポキシ)-tert-ブトキシホスホリルオキシ]プロピオン酸tert-ブチルエステル(318.0mg、0.460mmol)をCH3CN(16ml)およびH2O(4ml)の混合液に溶解させ、0℃で撹拌し、硝酸セリウムアンモニウム(629.1mg、1.147mmol)を加え、0℃で1時間攪拌した。H2O(20ml)を加えて反応をクエンチし、AcOEt(20ml)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=2:1→1:1)により精製し、表題の化合物を得た(238.2mg、0.407mmol、88%、褐色の油状物)。
(2S) -2-tert-Butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3- (4-methoxyphenoxy) propoxy) -tert-butoxyphosphoryloxy] propionic acid tert-Butyl ester (318.0 mg, 0.460 mmol) was dissolved in a mixture of CH 3 CN (16 ml) and H 2 O (4 ml), stirred at 0 ° C., cerium ammonium nitrate (629.1 mg, 1. 147 mmol) was added, and the mixture was stirred at 0 ° C. for 1 hour. The reaction was quenched by the addition of H 2 O (20 ml) and extracted with AcOEt (20 ml). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane: AcOEt = 2: 1 → 1: 1) to give the title compound (238.2 mg, 0.407 mmol, 88%, brown oil).
1H-NMR(CDCl3):δ=5.501 (1H, m), 4.363 (2H, m), 4.229 (1H, m), 3.987 (2H, m), 3.900 (1H, m), 3.629 (2H, m), 2.460 (1H, brs), 1.501-1.479 (18H, m), 1.450 (9H, s), 0.897 (9H, s), 0.110 (3H, s), 0.106 (3H, s)。
31P-NMR(CDCl3):δ=-4.977, -5.265。
HRMS (ESI, [M+Na]+):計算値:C25H52NNaO10PSi+: 608.2990;実測値:608.3013。 1 H-NMR (CDCl 3 ): δ = 5.501 (1H, m), 4.363 (2H, m), 4.229 (1H, m), 3.987 (2H, m), 3.900 (1H, m), 3.629 (2H, m), 2.460 (1H, brs), 1.501-1.479 (18H, m), 1.450 (9H, s), 0.897 (9H, s), 0.110 (3H, s), 0.106 (3H, s).
31 P-NMR (CDCl 3 ): δ = −4.977, −5.265.
HRMS (ESI, [M + Na] + ): Calculated: C 25 H 52 NNaO 10 PSi + : 608.2990; Found: 608.3013.
31P-NMR(CDCl3):δ=-4.977, -5.265。
HRMS (ESI, [M+Na]+):計算値:C25H52NNaO10PSi+: 608.2990;実測値:608.3013。 1 H-NMR (CDCl 3 ): δ = 5.501 (1H, m), 4.363 (2H, m), 4.229 (1H, m), 3.987 (2H, m), 3.900 (1H, m), 3.629 (2H, m), 2.460 (1H, brs), 1.501-1.479 (18H, m), 1.450 (9H, s), 0.897 (9H, s), 0.110 (3H, s), 0.106 (3H, s).
31 P-NMR (CDCl 3 ): δ = −4.977, −5.265.
HRMS (ESI, [M + Na] + ): Calculated: C 25 H 52 NNaO 10 PSi + : 608.2990; Found: 608.3013.
工程6:(2S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-ヒドロキシプロポキシ)-tert-ブトキシホスホリルオキシ]プロピオン酸tert-ブチルエステルの調製
Step 6: (2S) -2-tert-Butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3-hydroxypropoxy) -tert-butoxyphosphoryloxy] propionic acid tert- Preparation of butyl ester
(2S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-ヒドロキシプロポキシ)-tert-ブトキシホスホリルオキシ]プロピオン酸tert-ブチルエステル(75.4mg、0.129mmol)および3-(2-ヘプチルオキシフェニル)プロピオン酸(43.1mg、0.163mmol)をCH2Cl2(2ml)に溶解させ、4-ジメチルアミノピリジン(2.0mg、0.0166mmol)を加え、室温で1-エチル-3-(3-ジメチルアミノプロピル)カルボジイミド塩酸塩(33.0mg、0.172mmol)を加え、室温で15時間攪拌した。その後、MeOH(1ml)および1-エチル-3-(3-ジメチルアミノプロピル)カルボジイミド塩酸塩(30.1mg、0.157mmol)を加え、さらに2時間攪拌した。H2O(10ml)を加えて反応をクエンチし、CH2Cl2(10ml×3)で抽出した。合わせた有機層を食塩水で洗浄し、Na2SO4で乾燥させ、溶媒を留去した。残渣をカラムクロマトグラフィー(ヘキサン:AcOEt=4:1)により精製し、表題の化合物を得た(103.4mg、0.122mmol、95%、淡黄色の油状物)。
(2S) -2-tert-butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3-hydroxypropoxy) -tert-butoxyphosphoryloxy] propionic acid tert-butyl ester ( 75.4 mg, 0.129 mmol) and 3- (2-heptyloxyphenyl) propionic acid (43.1 mg, 0.163 mmol) were dissolved in CH 2 Cl 2 (2 ml) and 4-dimethylaminopyridine (2.0 mg 0.0166 mmol), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (33.0 mg, 0.172 mmol) was added at room temperature, and the mixture was stirred at room temperature for 15 hours. Thereafter, MeOH (1 ml) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (30.1 mg, 0.157 mmol) were added, and the mixture was further stirred for 2 hours. The reaction was quenched with H 2 O (10 ml) and extracted with CH 2 Cl 2 (10 ml × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and evaporated. The residue was purified by column chromatography (hexane: AcOEt = 4: 1) to give the title compound (103.4 mg, 0.122 mmol, 95%, pale yellow oil).
1H-NMR(CDCl3):δ=7.145 (2H, m), 6.829 (2H, m), 5.492 (1H, m), 4.350 (2H, m), 4.250-4.121 (2H, m), 4.004 (2H, m), 3.970-3.874 (4H, m), 2.940 (2H, t, J=7.78 Hz), 2.634 (2H, t, J=7.82 Hz), 1.793 (2H, m), 1.488 (9H, s), 1.469 (9H, s), 1.440 (9H, s), 1.360-1.301 (8H, m), 0.893 (3H, t, J=6.64 Hz), 0.876 (9H, s), 0.089 (3H, s), 0.076 (3H, s)。
13C-NMR(CDCl3):δ=173.03, 156.95, 155.35, 129.94, 128.78, 127.55, 120.18, 110.18, 82.64, 82.63, 79.91, 69.19, 69.10, 67.75, 67.49, 65.16, 34.01, 31.77, 29.82, 29.78, 29.74, 29.31, 29.03, 28.31, 27.95, 26.11, 26.08, 25.66, 22.60, 18.02, 14.08, -4.77, -4.83。
31P-NMR(CDCl3):δ=-5.632, -5.816。
HRMS (ESI, [M+Na]+):計算値:C41H74NNaO12PSi+: 854.4610;実測値:854.4610。 1 H-NMR (CDCl 3 ): δ = 7.145 (2H, m), 6.829 (2H, m), 5.492 (1H, m), 4.350 (2H, m), 4.250-4.121 (2H, m), 4.004 ( 2H, m), 3.970-3.874 (4H, m), 2.940 (2H, t, J = 7.78 Hz), 2.634 (2H, t, J = 7.82 Hz), 1.793 (2H, m), 1.488 (9H, s ), 1.469 (9H, s), 1.440 (9H, s), 1.360-1.301 (8H, m), 0.893 (3H, t, J = 6.64 Hz), 0.876 (9H, s), 0.089 (3H, s) , 0.076 (3H, s).
13 C-NMR (CDCl 3 ): δ = 173.03, 156.95, 155.35, 129.94, 128.78, 127.55, 120.18, 110.18, 82.64, 82.63, 79.91, 69.19, 69.10, 67.75, 67.49, 65.16, 34.01, 31.77, 29.82, 29.78 , 29.74, 29.31, 29.03, 28.31, 27.95, 26.11, 26.08, 25.66, 22.60, 18.02, 14.08, -4.77, -4.83.
31 P-NMR (CDCl 3 ): δ = −5.632, −5.816.
HRMS (ESI, [M + Na] + ): Calculated: C 41 H 74 NNaO 12 PSi + : 854.4610; Found: 854.4610.
13C-NMR(CDCl3):δ=173.03, 156.95, 155.35, 129.94, 128.78, 127.55, 120.18, 110.18, 82.64, 82.63, 79.91, 69.19, 69.10, 67.75, 67.49, 65.16, 34.01, 31.77, 29.82, 29.78, 29.74, 29.31, 29.03, 28.31, 27.95, 26.11, 26.08, 25.66, 22.60, 18.02, 14.08, -4.77, -4.83。
31P-NMR(CDCl3):δ=-5.632, -5.816。
HRMS (ESI, [M+Na]+):計算値:C41H74NNaO12PSi+: 854.4610;実測値:854.4610。 1 H-NMR (CDCl 3 ): δ = 7.145 (2H, m), 6.829 (2H, m), 5.492 (1H, m), 4.350 (2H, m), 4.250-4.121 (2H, m), 4.004 ( 2H, m), 3.970-3.874 (4H, m), 2.940 (2H, t, J = 7.78 Hz), 2.634 (2H, t, J = 7.82 Hz), 1.793 (2H, m), 1.488 (9H, s ), 1.469 (9H, s), 1.440 (9H, s), 1.360-1.301 (8H, m), 0.893 (3H, t, J = 6.64 Hz), 0.876 (9H, s), 0.089 (3H, s) , 0.076 (3H, s).
13 C-NMR (CDCl 3 ): δ = 173.03, 156.95, 155.35, 129.94, 128.78, 127.55, 120.18, 110.18, 82.64, 82.63, 79.91, 69.19, 69.10, 67.75, 67.49, 65.16, 34.01, 31.77, 29.82, 29.78 , 29.74, 29.31, 29.03, 28.31, 27.95, 26.11, 26.08, 25.66, 22.60, 18.02, 14.08, -4.77, -4.83.
31 P-NMR (CDCl 3 ): δ = −5.632, −5.816.
HRMS (ESI, [M + Na] + ): Calculated: C 41 H 74 NNaO 12 PSi + : 854.4610; Found: 854.4610.
工程7:(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(2-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸の調製
Step 7: Preparation of (2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (2-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid
(2S)-2-tert-ブチルカルボニルアミノ-3-[((2R)-2-(tert-ブチルジメチルシリルオキシ)-3-ヒドロキシプロポキシ)-tert-ブトキシホスホリルオキシ]プロピオン酸tert-ブチルエステル(83.6mg、0.0988mmol)を0℃でトリフルオロ酢酸(1.0ml)に溶解させ、室温で1時間攪拌した。反応混合物をトリフルオロ酢酸で希釈し、濃縮した。残渣をカラムクロマトグラフィー(CHCl3/MeOH/AcOH=8:1:1→6:1:3)により精製し、表題の化合物を酢酸塩として得た(38.5mg、0.0762mmol、77%、白色粉末)。得られた酢酸塩をトリフルオロ酢酸に溶解させ、溶媒を留去し、表題の化合物をトリフルオロ酢酸塩として得た(白色粉末)。
(2S) -2-tert-butylcarbonylamino-3-[((2R) -2- (tert-butyldimethylsilyloxy) -3-hydroxypropoxy) -tert-butoxyphosphoryloxy] propionic acid tert-butyl ester ( 83.6 mg, 0.0988 mmol) was dissolved in trifluoroacetic acid (1.0 ml) at 0 ° C. and stirred at room temperature for 1 hour. The reaction mixture was diluted with trifluoroacetic acid and concentrated. The residue was purified by column chromatography (CHCl 3 / MeOH / AcOH = 8: 1: 1 → 6: 1: 3) to give the title compound as an acetate salt (38.5 mg, 0.0762 mmol, 77%, White powder). The resulting acetate was dissolved in trifluoroacetic acid and the solvent was distilled off to give the title compound as a trifluoroacetate (white powder).
1H-NMR(CDCl3/TFA-d=4:1):δ=7.186 (1H, m), 7.087 (1H, m), 6.935-6.881 (2H, m), 4.667 (2H, m), 4.527 (1H, m), 4.297-4.173 (4H, m), 4.011 (2H, t, J=6.56 Hz), 2.967 (2H, m), 2.793 (2H, m), 1.790 (2H, m), 1.438 (2H, m), 1.367-1.268 (7H,m) 0.880 (3H, t, J=6.52 Hz)。
31P-NMR(CDCl3):δ=-1.374。
融点:134.2-135.0 ℃。
元素分析:計算値:C22H36NO10P?0.6CF3COOH: C, 48.55; H, 6.43; N, 2.44;実測値:C, 48.45; H, 6.76; N, 2.56。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 7.186 (1H, m), 7.087 (1H, m), 6.935-6.881 (2H, m), 4.667 (2H, m), 4.527 (1H, m), 4.297-4.173 (4H, m), 4.011 (2H, t, J = 6.56 Hz), 2.967 (2H, m), 2.793 (2H, m), 1.790 (2H, m), 1.438 ( 2H, m), 1.367-1.268 (7H, m) 0.880 (3H, t, J = 6.52 Hz).
31 P-NMR (CDCl 3 ): δ = -1.374.
Melting point: 134.2-135.0 ° C.
? Calcd: C 22 H 36 NO 10 P 0.6CF 3 COOH: C, 48.55; H, 6.43; N, 2.44; Found: C, 48.45; H, 6.76 ; N, 2.56.
31P-NMR(CDCl3):δ=-1.374。
融点:134.2-135.0 ℃。
元素分析:計算値:C22H36NO10P?0.6CF3COOH: C, 48.55; H, 6.43; N, 2.44;実測値:C, 48.45; H, 6.76; N, 2.56。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 7.186 (1H, m), 7.087 (1H, m), 6.935-6.881 (2H, m), 4.667 (2H, m), 4.527 (1H, m), 4.297-4.173 (4H, m), 4.011 (2H, t, J = 6.56 Hz), 2.967 (2H, m), 2.793 (2H, m), 1.790 (2H, m), 1.438 ( 2H, m), 1.367-1.268 (7H, m) 0.880 (3H, t, J = 6.52 Hz).
31 P-NMR (CDCl 3 ): δ = -1.374.
Melting point: 134.2-135.0 ° C.
? Calcd: C 22 H 36 NO 10 P 0.6
上記実施例と同様の手法により、以下の化合物を調製した。
The following compounds were prepared by the same method as in the above examples.
[実施例31](2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(3-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 31] (2S) -2-Amino-3-[((2R) -2-hydroxy-3- {3- (3-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ=7.248 (1H, m), 6.865-6.822 (3H, m), 4.692-4.291 (6H, m), 4.115 (2H, m), 3.257 (2H, m), 2.944 (2H, m), 2.789 (2H, m), 1.787 (2H, t, J=6.92 Hz), 1.426-1.278 (8H, m), 0.882 (3H, t, J=6.48 Hz)。
31P-NMR(CDCl3):δ=-1.298。
融点:158.4-159.5 ℃。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 7.248 (1H, m), 6.865-6.822 (3H, m), 4.692-4.291 (6H, m), 4.115 (2H, m) , 3.257 (2H, m), 2.944 (2H, m), 2.789 (2H, m), 1.787 (2H, t, J = 6.92 Hz), 1.426-1.278 (8H, m), 0.882 (3H, t, J = 6.48 Hz).
31 P-NMR (CDCl 3 ): δ = -1.298.
Melting point: 158.4-159.5 ° C.
31P-NMR(CDCl3):δ=-1.298。
融点:158.4-159.5 ℃。 1 H-NMR (CDCl 3 / TFA-d = 4: 1 ): δ = 7.248 (1H, m), 6.865-6.822 (3H, m), 4.692-4.291 (6H, m), 4.115 (2H, m) , 3.257 (2H, m), 2.944 (2H, m), 2.789 (2H, m), 1.787 (2H, t, J = 6.92 Hz), 1.426-1.278 (8H, m), 0.882 (3H, t, J = 6.48 Hz).
31 P-NMR (CDCl 3 ): δ = -1.298.
Melting point: 158.4-159.5 ° C.
[実施例32](2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(4-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 32] (2S) -2-Amino-3-[((2R) -2-hydroxy-3- {3- (4-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ=7.103 (2H, d, J=8.04 Hz), 6.912 (2H, d, J=7.98 Hz), 4.665 (2H, m), 4.296 (1H, m), 4.207 (3H, m), 4.088 (2H, t, J=6.82 Hz), 3.835-3.605 (2H, m), 2.909 (2H, m), 2.748 (2H, m), 1.776 (2H, m), 1.419-1.316 (9H, m), 0.881 (3H, t, J=6.72 Hz)。
31P-NMR(CDCl3):δ=-1.380。
融点:163.7-164.6 ℃。
元素分析:計算値:C22H36NO10P?0.4CF3COOH: C, 49.69; H, 6.66; N, 2.54;実測値:C, 49.42; H, 6.95; N, 2.78。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 7.103 (2H, d, J = 8.04 Hz), 6.912 (2H, d, J = 7.98 Hz), 4.665 (2H, m), 4.296 (1H, m), 4.207 (3H, m), 4.088 (2H, t, J = 6.82 Hz), 3.835-3.605 (2H, m), 2.909 (2H, m), 2.748 (2H, m), 1.776 (2H, m), 1.419-1.316 (9H, m), 0.881 (3H, t, J = 6.72 Hz).
31 P-NMR (CDCl 3 ): δ = -1.380.
Melting point: 163.7-164.6 ° C.
? Calcd: C 22 H 36 NO 10 P 0.4CF 3 COOH: C, 49.69; H, 6.66; N, 2.54; Found: C, 49.42; H, 6.95 ; N, 2.78.
31P-NMR(CDCl3):δ=-1.380。
融点:163.7-164.6 ℃。
元素分析:計算値:C22H36NO10P?0.4CF3COOH: C, 49.69; H, 6.66; N, 2.54;実測値:C, 49.42; H, 6.95; N, 2.78。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 7.103 (2H, d, J = 8.04 Hz), 6.912 (2H, d, J = 7.98 Hz), 4.665 (2H, m), 4.296 (1H, m), 4.207 (3H, m), 4.088 (2H, t, J = 6.82 Hz), 3.835-3.605 (2H, m), 2.909 (2H, m), 2.748 (2H, m), 1.776 (2H, m), 1.419-1.316 (9H, m), 0.881 (3H, t, J = 6.72 Hz).
31 P-NMR (CDCl 3 ): δ = -1.380.
Melting point: 163.7-164.6 ° C.
? Calcd: C 22 H 36 NO 10 P 0.4
[実施例33](2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(4-ベンジルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 33] (2S) -2-Amino-3-[((2R) -2-hydroxy-3- {3- (4-benzyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid
1H-NMR(CDCl3/TFA-d=4:1):δ=7.341-7.291 (4H, m), 7.237-7.165 (1H, m), 7.101 (2H, d, J=8.08 Hz), 6.952 (2H, d, J=8.00 Hz), 5.120 (2H, s), 4.654 (2H, m), 4.492 (1H, m), 4.301-4.198 (4H, m), 3.833-3.779 (1H, m), 2.902 (2H, m), 2.744 (2H, m), 1.271 (1H, s)。
31P-NMR(CDCl3):δ=-1.366。
融点:131.0-132.1 ℃。
元素分析:C22H28NO10P?0.8CF3COOH?1.2H2O: C, 46.45; H, 5.15; N, 2.30;実測値:C, 46.55; H, 5.36; N, 2.63。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 7.341-7.291 (4H, m), 7.237-7.165 (1H, m), 7.101 (2H, d, J = 8.08 Hz), 6.952 (2H, d, J = 8.00 Hz), 5.120 (2H, s), 4.654 (2H, m), 4.492 (1H, m), 4.301-4.198 (4H, m), 3.833-3.779 (1H, m), 2.902 (2H, m), 2.744 (2H, m), 1.271 (1H, s).
31 P-NMR (CDCl 3 ): δ = -1.366.
Melting point: 131.0-132.1 ° C.
?? Elemental analysis: C 22 H 28 NO 10 P 0.8CF 3 COOH 1.2H 2 O: C, 46.45; H, 5.15; N, 2.30; Found: C, 46.55; H, 5.36 ; N, 2.63.
31P-NMR(CDCl3):δ=-1.366。
融点:131.0-132.1 ℃。
元素分析:C22H28NO10P?0.8CF3COOH?1.2H2O: C, 46.45; H, 5.15; N, 2.30;実測値:C, 46.55; H, 5.36; N, 2.63。 1 H-NMR (CDCl 3 / TFA-d = 4: 1): δ = 7.341-7.291 (4H, m), 7.237-7.165 (1H, m), 7.101 (2H, d, J = 8.08 Hz), 6.952 (2H, d, J = 8.00 Hz), 5.120 (2H, s), 4.654 (2H, m), 4.492 (1H, m), 4.301-4.198 (4H, m), 3.833-3.779 (1H, m), 2.902 (2H, m), 2.744 (2H, m), 1.271 (1H, s).
31 P-NMR (CDCl 3 ): δ = -1.366.
Melting point: 131.0-132.1 ° C.
?? Elemental analysis: C 22 H 28 NO 10 P 0.8
以下の化合物は、Avanti Polar Lipids(858143C)より購入したものを使用した。
The following compounds were purchased from Avanti® Polar Lipids (858143C).
[実施例34](2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸(LPS(18:1))
Example 34 (2S) -2-amino-3-[((2R) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid (LPS (18: 1))
以下の化合物は、特開2007-284402に記載の方法により調製した。
The following compounds were prepared by the method described in JP-A-2007-284402.
[実施例35](2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
Example 35 (2S) -2-Amino-3-[((2R) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
[実施例36](2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
[Example 36] (2S) -2-Amino-3-[((2R) -2-hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid
本発明に使用される化合物、または本発明の化合物の合成方法を以下のスキームに示す。
The compound used in the present invention or the synthesis method of the compound of the present invention is shown in the following scheme.
[試験例1]TGFα切断アッセイによるアゴニスト活性の評価
[Test Example 1] Evaluation of agonist activity by TGFα cleavage assay
工程1:P2Y10およびGPR174遺伝子のクローニング
HUVEC細胞、マウス尾ゲノム、ラット尾ゲノムをテンプレートとして、PCR法により、ヒト、マウス及びラットのP2Y10とGPR174遺伝子の全長cDNAを取得した。ヒト、マウスおよびラットのP2Y10とGPR174遺伝子は配列番号1~6で表される塩基配列からなるオリゴヌクレオチドをフォワードプライマーとして、配列番号7~12で表される塩基配列からなるオリゴヌクレオチドをリバースプライマーとして用いた。 Step 1: Cloning of P2Y10 and GPR174 genes Full-length cDNAs of human, mouse, and rat P2Y10 and GPR174 genes were obtained by PCR using HUVEC cells, mouse tail genome, and rat tail genome as templates. For human, mouse and rat P2Y10 and GPR174 genes, the oligonucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 1-6 is used as a forward primer, and the oligonucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 7-12 is used as a reverse primer Using.
HUVEC細胞、マウス尾ゲノム、ラット尾ゲノムをテンプレートとして、PCR法により、ヒト、マウス及びラットのP2Y10とGPR174遺伝子の全長cDNAを取得した。ヒト、マウスおよびラットのP2Y10とGPR174遺伝子は配列番号1~6で表される塩基配列からなるオリゴヌクレオチドをフォワードプライマーとして、配列番号7~12で表される塩基配列からなるオリゴヌクレオチドをリバースプライマーとして用いた。 Step 1: Cloning of P2Y10 and GPR174 genes Full-length cDNAs of human, mouse, and rat P2Y10 and GPR174 genes were obtained by PCR using HUVEC cells, mouse tail genome, and rat tail genome as templates. For human, mouse and rat P2Y10 and GPR174 genes, the oligonucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 1-6 is used as a forward primer, and the oligonucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 7-12 is used as a reverse primer Using.
なお、前記フォワードプライマーの各々の5 '末端には、制限酵素Kpn I認識部位を含む塩基配列がそれぞれ付加されている。また、リバースプライマーのヒトP2Y10の3 '末端には、制限酵素Sma I認識部位を含む塩基配列が付加されており、ヒトGPR174、マウス及びラットのP2Y10とGPR174の3 '末端には、制限酵素EcoR V認識部位を含む塩基配列がそれぞれ付加されている。PCRは、PrimeSTAR HS(#R010Q, TAKARA)を用いて、98℃(10秒間)/ 55℃(15秒間)/ 72℃(1分間)からなるサイクルを40回繰り返した。その結果、約1kbpのDNA断片が増幅された。Mighty Mix(タカラバイオ社)、HiSpeed Plasmid MidiKit (QIAGEN社)の各キットを使用して、このDNA断片をKpn I、Sma I及びEcoR Vで消化した後、プラスミドpCAGGSのマルチクローニングサイトに挿入することにより、それぞれのプラスミドを得た。ヒト、マウス及びラットのP2Y10とGPR174遺伝子の塩基配列は、株式会社 BEXに委託し、ダイターミネーター法により決定した。P2Y10遺伝子の塩基配列は、配列番号13~15で表される塩基配列のとおりであり、GPR174遺伝子の塩基配列は配列番号16~18で表される塩基配列のとおりであった。
A base sequence containing a restriction enzyme KpnKI recognition site is added to the 5 ′ end of each of the forward primers. In addition, a base sequence including a restriction enzyme Sma I recognition site is added to the 3 ′ ′ end of human P2Y10 of the reverse primer, and restriction enzyme EcoR is added to the 3 ′ ′ end of human GPR174, mouse and rat P2Y10 and GPR174. Each nucleotide sequence including a V recognition site is added. For PCR, PrimeSTARPrimHS (# R010Q, 010TAKARA) was used, and a cycle consisting of 98 ° C. (10 seconds) / 55 ° C. (15 seconds) / 72 ° C. (1 minute) was repeated 40 times. As a result, a DNA fragment of about 1 kbp was amplified. Using Mighty 断 片 Mix (Takara Bio) and HiSpeed 及 び Plasmid MidiKit (QIAGEN) kits, this DNA fragment should be digested with Kpn I, Sma I and EcoR V and then inserted into the multicloning site of plasmid pCAGGS. Thus, each plasmid was obtained. The nucleotide sequences of human, mouse and rat P2Y10 and GPR174 genes were consigned to Sakai BEX and determined by the dye terminator method. The base sequence of the P2Y10 gene was as shown in SEQ ID NOs: 13 to 15, and the base sequence of the GPR174 gene was as shown in SEQ ID NOs: 16 to 18.
配列番号13~15で表される塩基配列は、ヒト、マウス、およびラットでそれぞれ984塩基、987塩基、987塩基のオープンリーディングフレーム(O R F)を有しており、このO R Fから予測されるアミノ酸配列(それぞれ328アミノ酸、329アミノ酸、329アミノ酸)は、配列番号19~21で表されるアミノ酸配列のとおりであった。また、配列番号16~18で表される塩基配列は、ヒト、マウス、およびラットでそれぞれ1002塩基、1008塩基、1008塩基のORFを有しており、このORFから予測されるアミノ酸配列(それぞれ334アミノ酸、336アミノ酸、336アミノ酸)は、配列番号22~24で表されるアミノ酸配列のとおりであった。
The base sequences represented by SEQ ID NOs: 13 to 15 have open reading frames (O R F) of 984 bases, 987 bases, and 987 bases in human, mouse, and rat, respectively, and are predicted from this O R F. The amino acid sequences (328 amino acids, 329 amino acids, and 329 amino acids) were as shown in SEQ ID NOs: 19 to 21, respectively. Further, the base sequences represented by SEQ ID NOs: 16 to 18 have ORFs of 1002 bases, 1008 bases and 1008 bases in human, mouse and rat, respectively, and amino acid sequences predicted from these ORFs (each 334 Amino acids, 336 amino acids, 336 amino acids) were as shown in the amino acid sequences represented by SEQ ID NOs: 22 to 24.
工程2:TGFα切断アッセイによるアゴニスト活性の評価
HEK293細胞を2.0x105cells/mLとなるように10%ウシ胎仔血清(FCS)含有のダルベッコ変法イーグル培地(DMEM)で懸濁し、60mmディッシュに4mLずつ播種した。5%CO2存在下で24時間培養後、LipofectAMINE 2000 (Invitrogen社)を用いて各種発現ベクターのトランスフェクションを行った。60mmディッシュあたり、AP標識TGFαのプラスミドベクターを1μg、マウスのP2Y10またはGPR174のプラスミドベクター1μgを用いた。なお、AP標識TGFαは膜結合型pro-TGFαのN末端側にhuman placental alkaline phosphataseが融合したタンパクであり、AP標識TGFαのプラスミドベクターは、Tokumaru et al., J Cell Biol 151, 209-220 (2000)の記載に基づいて作成することができる。トランスフェクション24時間後、トリプシン/EDTAを用いて細胞を剥がし、2.0x104cells/wellとなるように培養液に再懸濁し、96-wellプレートに播種した。化合物で刺激する場合は再懸濁液にハンクス平衡塩溶液(HBSS、5mM HEPES含有)を用いた。30分間静置後、各種濃度の被検化合物を添加し、5%CO2存在下でさらに1時間静置した。96-wellプレートを遠心 (190 x g , 3分)し、上清80μLを別の96-wellプレートに移した。10 mM p-ニトロフェニルホスフェート(p-NPP)を含有した反応バッファー(40 mM Tris-HCl (pH 9.5))を上清及び細胞に80μL加えた。OD405をマイクロプレートリーダーで測定し(バックグラウンド)、37℃で1時間加温した後にOD405を再度測定した。各wellについて、AP活性は2回目の吸光度からバックグラウンドを引いた値を以下の式に当てはめて算出した。 Step 2: Evaluation of agonist activity by TGFα cleavage assay HEK293 cells are suspended in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal calf serum (FCS) at 2.0x10 5 cells / mL, and 4 mL in a 60 mm dish. Seeded one by one. After culturing for 24 hours in the presence of 5% CO 2 , various expression vectors were transfected using LipofectAMINE 2000 (Invitrogen). 1 μg of AP-tagged TGFα plasmid vector and 1 μg of mouse P2Y10 or GPR174 plasmid vector were used per 60 mm dish. AP-labeled TGFα is a protein in which human placental alkaline phosphatase is fused to the N-terminal side of membrane-bound pro-TGFα, and the AP-labeled TGFα plasmid vector is Tokumaru et al., J Cell Biol 151, 209-220 ( 2000). Twenty-four hours after transfection, the cells were detached using trypsin / EDTA, resuspended in a culture solution to 2.0 × 10 4 cells / well, and seeded in a 96-well plate. When stimulating with a compound, Hanks balanced salt solution (HBSS, containing 5 mM HEPES) was used for resuspension. After standing for 30 minutes, various concentrations of test compounds were added, and the mixture was further allowed to stand for 1 hour in the presence of 5% CO 2 . The 96-well plate was centrifuged (190 × g, 3 minutes), and 80 μL of the supernatant was transferred to another 96-well plate. 80 μL of a reaction buffer (40 mM Tris-HCl (pH 9.5)) containing 10 mM p-nitrophenyl phosphate (p-NPP) was added to the supernatant and cells. OD405 was measured with a microplate reader (background), heated at 37 ° C. for 1 hour, and then measured again. For each well, the AP activity was calculated by applying a value obtained by subtracting the background from the second absorbance to the following equation.
HEK293細胞を2.0x105cells/mLとなるように10%ウシ胎仔血清(FCS)含有のダルベッコ変法イーグル培地(DMEM)で懸濁し、60mmディッシュに4mLずつ播種した。5%CO2存在下で24時間培養後、LipofectAMINE 2000 (Invitrogen社)を用いて各種発現ベクターのトランスフェクションを行った。60mmディッシュあたり、AP標識TGFαのプラスミドベクターを1μg、マウスのP2Y10またはGPR174のプラスミドベクター1μgを用いた。なお、AP標識TGFαは膜結合型pro-TGFαのN末端側にhuman placental alkaline phosphataseが融合したタンパクであり、AP標識TGFαのプラスミドベクターは、Tokumaru et al., J Cell Biol 151, 209-220 (2000)の記載に基づいて作成することができる。トランスフェクション24時間後、トリプシン/EDTAを用いて細胞を剥がし、2.0x104cells/wellとなるように培養液に再懸濁し、96-wellプレートに播種した。化合物で刺激する場合は再懸濁液にハンクス平衡塩溶液(HBSS、5mM HEPES含有)を用いた。30分間静置後、各種濃度の被検化合物を添加し、5%CO2存在下でさらに1時間静置した。96-wellプレートを遠心 (190 x g , 3分)し、上清80μLを別の96-wellプレートに移した。10 mM p-ニトロフェニルホスフェート(p-NPP)を含有した反応バッファー(40 mM Tris-HCl (pH 9.5))を上清及び細胞に80μL加えた。OD405をマイクロプレートリーダーで測定し(バックグラウンド)、37℃で1時間加温した後にOD405を再度測定した。各wellについて、AP活性は2回目の吸光度からバックグラウンドを引いた値を以下の式に当てはめて算出した。 Step 2: Evaluation of agonist activity by TGFα cleavage assay HEK293 cells are suspended in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal calf serum (FCS) at 2.0x10 5 cells / mL, and 4 mL in a 60 mm dish. Seeded one by one. After culturing for 24 hours in the presence of 5% CO 2 , various expression vectors were transfected using LipofectAMINE 2000 (Invitrogen). 1 μg of AP-tagged TGFα plasmid vector and 1 μg of mouse P2Y10 or GPR174 plasmid vector were used per 60 mm dish. AP-labeled TGFα is a protein in which human placental alkaline phosphatase is fused to the N-terminal side of membrane-bound pro-TGFα, and the AP-labeled TGFα plasmid vector is Tokumaru et al., J Cell Biol 151, 209-220 ( 2000). Twenty-four hours after transfection, the cells were detached using trypsin / EDTA, resuspended in a culture solution to 2.0 × 10 4 cells / well, and seeded in a 96-well plate. When stimulating with a compound, Hanks balanced salt solution (HBSS, containing 5 mM HEPES) was used for resuspension. After standing for 30 minutes, various concentrations of test compounds were added, and the mixture was further allowed to stand for 1 hour in the presence of 5% CO 2 . The 96-well plate was centrifuged (190 × g, 3 minutes), and 80 μL of the supernatant was transferred to another 96-well plate. 80 μL of a reaction buffer (40 mM Tris-HCl (pH 9.5)) containing 10 mM p-nitrophenyl phosphate (p-NPP) was added to the supernatant and cells. OD405 was measured with a microplate reader (background), heated at 37 ° C. for 1 hour, and then measured again. For each well, the AP activity was calculated by applying a value obtained by subtracting the background from the second absorbance to the following equation.
さらに、無刺激群のAP活性(%)を差し引いた値を縦軸に用いてグラフを作成した。実施例34(LPS(18:1))、実施例1(deoxy-LPS(18:1))および実施例19(LPalloT(18:1))についての試験結果のグラフを図1および2に示す。図1および2のグラフより、deoxy-LPS(18:1)はP2Y10に、LPalloT(18:1)はGPR174にそれぞれ選択性を有するアゴニストであることが確認された。
Furthermore, a graph was created using the value obtained by subtracting the AP activity (%) of the unstimulated group on the vertical axis. Graphs of test results for Example 34 (LPS (18: 1)), Example 1 (deoxy-LPS (18: 1)) and Example 19 (LPalloT (18: 1)) are shown in FIGS. . From the graphs of FIGS. 1 and 2, it was confirmed that deoxy-LPS (18: 1) is an agonist having selectivity for P2Y10, and LPalloT (18: 1) is a selectivity for GPR174.
また各試験化合物について算出した最大反応値(Emax)を実施例34(LPS(18:1))のEmaxと比較して、各化合物のアゴニスト活性を次のように表示した: 0-40%:-、40-60%:+、60-80%:++、80-100%:+++、試験結果なし:ND。結果を以下の表1および2に示す。
Further, the maximum response value (Emax) calculated for each test compound was compared with Emax of Example 34 (LPS (18: 1)), and the agonist activity of each compound was expressed as follows: 0-40%: -, 40-60%: +, 60-80%: ++, 80-100%: +++, no test result: ND. The results are shown in Tables 1 and 2 below.
[試験例2]全身性エリテマトーデス自然発症マウスにおける評価
MRL-lpr/lprマウス(以下、MRL/lprマウスとも称する、8~9 週齢,♀)は日本SLC株式会社より購入した。実施例34の化合物(LPS(18:1)、Avanti Polar Lipidsより購入、858143C)をマイクロシリンジでシリコンコーティング済みのガラス管(旭テクノグラス,9831-1207)に取り、窒素で乾固した後、0.01%(w/v)BSA-PBSを加えボルテックスを行い bath sonicator で2分間ソニケーションを行い10mM LPSに調製した。 [Test Example 2] Evaluation in spontaneous systemic lupus erythematosus mice MRL-lpr / lpr mice (hereinafter also referred to as MRL / lpr mice, 8-9 weeks old, rabbit) were purchased from Japan SLC Co., Ltd. The compound of Example 34 (LPS (18: 1), purchased from Avanti Polar Lipids, 858143C) was taken into a silicon-coated glass tube (Asahi Techno Glass, 9831-1207) with a microsyringe and dried to nitrogen, 0.01% (w / v) BSA-PBS was added, vortexed, and sonicated with a bath sonicator for 2 minutes to prepare 10 mM LPS.
MRL-lpr/lprマウス(以下、MRL/lprマウスとも称する、8~9 週齢,♀)は日本SLC株式会社より購入した。実施例34の化合物(LPS(18:1)、Avanti Polar Lipidsより購入、858143C)をマイクロシリンジでシリコンコーティング済みのガラス管(旭テクノグラス,9831-1207)に取り、窒素で乾固した後、0.01%(w/v)BSA-PBSを加えボルテックスを行い bath sonicator で2分間ソニケーションを行い10mM LPSに調製した。 [Test Example 2] Evaluation in spontaneous systemic lupus erythematosus mice MRL-lpr / lpr mice (hereinafter also referred to as MRL / lpr mice, 8-9 weeks old, rabbit) were purchased from Japan SLC Co., Ltd. The compound of Example 34 (LPS (18: 1), purchased from Avanti Polar Lipids, 858143C) was taken into a silicon-coated glass tube (Asahi Techno Glass, 9831-1207) with a microsyringe and dried to nitrogen, 0.01% (w / v) BSA-PBS was added, vortexed, and sonicated with a bath sonicator for 2 minutes to prepare 10 mM LPS.
小型浸透圧ポンプ(Alzet社, model 2006)を生理食塩水(大塚生食注)に浸し、37℃で60時間インキュベートを行った。その後、1mL シリンジ(TERUMO,SS-01T)と付属の27Gの針を用いて各浸透圧ポンプにLPS溶液及び0.01%(w/v)BSA-PBSを200μLずつ充填した。Balb/cマウスにネンブタール麻酔薬を100mg/kgとなるように投与後、小型浸透圧ポンプをマウスの腹腔内に縦に埋め込み、腹膜、腹壁を縫合した。
A small osmotic pump (Alzet, model 2006) was soaked in physiological saline (Otsuka raw food injection) and incubated at 37 ° C for 60 hours. Thereafter, each osmotic pump was filled with 200 μL of LPS solution and 0.01% (w / v) BSA-PBS using a 1 mL syringe (TERUMO, SS-01T) and the attached 27G needle. After administration of Nembutal anesthetic to Balb / c mice at 100 mg / kg, a small osmotic pump was implanted vertically into the abdominal cavity of the mouse, and the peritoneum and abdominal wall were sutured.
以上の手順により、MRL/lprマウス(♀、9週齢)に実施例34の化合物(LPS(18:1), 10 mM)を充填した小型浸透圧ポンプ(排出速度0.15 μL/hr, 42日間)を腹腔に埋め込むことにより、LPS(18:1)を持続的に供給した。17週齢にて、採血および臓器の回収を行った。血漿を用いて全自動血球装置(VetScan HM2)による血球細胞数の測定を行い、血清を用いてELISAによる自己抗体量の測定を行い、さらにリンパ組織は水分除去後、電子天秤で各臓器の重量を測定した。
By the above procedure, MRL / lpr mice (♀, 9 weeks old) were filled with the compound of Example 34 (LPS (18: 1), 10 mM) (small discharge rate 0.15 μL / hr, 42 days) ) Was implanted into the abdominal cavity to provide a continuous supply of LPS (18: 1). At 17 weeks of age, blood was collected and organs were collected. Using blood plasma, the number of blood cells is measured with a fully automated blood cell device (VetScan HM2), the amount of autoantibodies is measured by ELISA using serum, and the lymph tissue is dehydrated, and the weight of each organ is measured with an electronic balance. Was measured.
自己抗体量の測定(抗dsDNA抗体の検出)は以下の手順により行った。96 well アッセイプレートにポリ-L-リシン溶液(50μL/well)をまき、室温で8時間静置し well底をコーティングした。アスピレーターで溶液をよく取り除いた後、1ng/μL DNA溶液を100μl/well加え、37℃で8時間インキュベートし、プレートに結合させた。DNA溶液を取り除いた後、Blocking bufferを180μL/well加え、室温で2時間blockingを行った。Blocking bufferをよく除きAssay bufferで希釈した血清サンプルを50μL/well加え、室温で2時間静置した。その後、Skan Washer 400(Molecular Devices)を用いてTTBSで3回 washし、Assay bufferで2000倍に希釈したbiotinylated anti-mouse IgG(H+L)(Vector,BA2001)を50μL/well加え2時間反応させた。さらに、TTBSで3回 washし、Assay bufferで2000倍希釈したStreptavidin-HRP(Invitrogen,434323)を50μL/well加えて1時間反応させ、最後にTTBSで6回 wash後、TMB発色を10~15分間行い、1M H3PO4 で反応停止後、450nm での吸光度をマイクロプレートリーダー(VERSAmax, Molecular Devices)で測定した。
Measurement of the amount of autoantibodies (detection of anti-dsDNA antibody) was performed by the following procedure. A 96-well assay plate was coated with a poly-L-lysine solution (50 μL / well) and allowed to stand at room temperature for 8 hours to coat the well bottom. After thoroughly removing the solution with an aspirator, 100 μl / well of 1 ng / μL DNA solution was added and incubated at 37 ° C. for 8 hours to bind to the plate. After removing the DNA solution, 180 μL / well of blocking buffer was added, and blocking was performed at room temperature for 2 hours. After removing the blocking buffer well, a serum sample diluted with assay buffer was added at 50 μL / well and allowed to stand at room temperature for 2 hours. Then wash 3 times with TTBS using Skan Washer 400 (Molecular Devices), add 50 μL / well of biotinylated anti-mouse IgG (H + L) (Vector, BA2001) diluted 2000 times with Assay buffer, and react for 2 hours. I let you. Further, wash 3 times with TTBS, add 50 μL / well of Streptavidin-HRP (Invitrogen, 434323) diluted 2000-fold with Assay buffer, react for 1 hour, finally wash 6 times with TTBS, and then develop TMB coloration 10-15 After stopping the reaction with 1M H 3 PO 4 , the absorbance at 450 nm was measured with a microplate reader (VERSAmax, Molecular Devices).
また、腎臓のパラフィン切片を作製し、anti-mouse IgG抗体を用いて免疫複合体の染色を行った。パラフィン切片をXylen(3回)、100%(3回)、90%、70%、50% EtOHにそれぞれ3分間ずつ順次浸し脱パラフィンを行った後、流水で5分間洗浄した。その後、3% H2O2 にスライドを30分間浸し組織の内在性のペルオキシダーゼの不活化を行い、再度流水で5分間洗浄した。圧力鍋を用い、1mM EDTA中にスライドガラスを浸し抗原の賦活化を行った後、Avidin/Biotin Blocking kit(VECTOR, SP-2001)を用いて内在のビオチンのブロッキングを行なった。その後、スライドに 1% BSA-PBS を添加し、湿箱内にて室温で2時間 Blockingを行った。次いで、スライドガラスに 1% BSA-PBS で2000倍に希釈したbiotinylated anti-mouse IgG(H+L)を添加して室箱内にて4℃で一晩反応させた。反応終了後、PBSで5分間ずつ3回ずつ洗浄した。その後、ABC試薬を添加して室温で30分間反応させた後、PBSで5分間ずつ3回ずつ洗浄した。発色はtyramide working solutionを30μL/specimen添加し室温で5~10分間反応させた。その後、PBSで洗浄を行いDAPIを添加し5分間核染色を行った後、余分な液をPBSで洗い落とし、封入剤Aqueous Mounting Medium, PermaFluor(Thermo,TA-030-FM)を用いて封入した。
In addition, kidney paraffin sections were prepared and immunocomplexes were stained with anti-mouse IgG antibodies. Paraffin sections were deparaffinized by sequentially immersing them in Xylen (3 times), 100% (3 times), 90%, 70%, and 50% EtOH for 3 minutes each, and then washed with running water for 5 minutes. Thereafter, the slide was immersed in 3% H 2 O 2 for 30 minutes to inactivate the tissue's endogenous peroxidase, and washed again with running water for 5 minutes. Using a pressure cooker, the slide glass was immersed in 1 mM EDTA to activate the antigen, and then endogenous biotin was blocked using an Avidin / Biotin Blocking kit (VECTOR, SP-2001). Thereafter, 1% BSA-PBS was added to the slide, and blocking was performed in a wet box at room temperature for 2 hours. Subsequently, biotinylated anti-mouse IgG (H + L) diluted 2000-fold with 1% BSA-PBS was added to the slide glass, and reacted overnight at 4 ° C. in a chamber box. After completion of the reaction, the cells were washed 3 times for 5 minutes each with PBS. Thereafter, ABC reagent was added and reacted at room temperature for 30 minutes, and then washed with PBS three times for 5 minutes each. For color development, 30 μL / specimen of tyramide working solution was added and reacted at room temperature for 5 to 10 minutes. Thereafter, the plate was washed with PBS, DAPI was added and nuclear staining was performed for 5 minutes, and then the excess solution was washed off with PBS and encapsulated using an encapsulating agent, Aqueous Mounting Medium, PermaFluor (Thermo, TA-030-FM).
上記の試験結果を図3~8に示す。試験結果より、LPS(18:1)は血中白血球数の減少(図3)、自己抗体産生の抑制(図4)、リンパ組織である脾臓および腸間膜リンパ節の腫脹の抑制(図5および6)、腎臓への免疫複合体の沈着抑制(図7および8)など、SLE様自己免疫疾患症状の発症抑制作用を有することが明らかとなった。
The above test results are shown in FIGS. From the test results, LPS (18: 1) reduced blood white blood cell count (Fig. 3), suppressed autoantibody production (Fig. 4), and suppressed swelling of lymphoid spleen and mesenteric lymph nodes (Fig. 5). And 6), it has been clarified that it has an inhibitory effect on the onset of SLE-like autoimmune disease symptoms such as suppression of immune complex deposition in the kidney (FIGS. 7 and 8).
[試験例3]ヒト自己免疫疾患肝炎モデルマウスにおける評価
野生型Balb/cマウス(♂、9~10週齢)に試験化合物(1.5 mM)を充填した小型浸透圧ポンプ(排出速度1.0μL/hr, 7日間)を腹腔に埋め込むことにより、LPS(18:1)、実施例1の化合物(deoxy-LPS(18:1))または実施例19(LPalloT(18:1))を持続的に供給した。小型浸透圧ポンプ(Alzet社, model 2001)は生理食塩水(大塚生食注)に浸し、37℃で4時間インキュベートしてから用い、それ以外は試験例2と同様の手順で小型浸透圧ポンプをマウスの腹腔に埋め込んだ。4日後に12時間断食し、コンカナバリンA(ConA)を20 mg/kgで尾静注した。一定時間後、採血を行い、血清を調製した。PBSにて灌流後、肝臓を摘出し、パラフィンブロック、切片の作製を行った。トランスアミナーゼCIIテストワコーを用いて血清中のアラニントランスアミナーゼ(ALT)を測定した。パラフィン切片はヘマトキシリンエオジン染色を行い、光学顕微鏡を用いて観察した。 [Test Example 3] Evaluation in a human autoimmune disease hepatitis model mouse A small osmotic pump (discharge rate 1.0 μL / hr) filled with a test compound (1.5 mM) in a wild-type Balb / c mouse (♂, 9 to 10 weeks old) , 7 days) continuously implanted LPS (18: 1), compound of Example 1 (deoxy-LPS (18: 1)) or Example 19 (LPalloT (18: 1)) did. The small osmotic pump (Alzet, model 2001) is immersed in physiological saline (Otsuka raw food injection) and incubated at 37 ° C for 4 hours before use. Otherwise, the small osmotic pump is used in the same procedure as in Test Example 2. Implanted in the abdominal cavity of mice. After 4 days, the animals were fasted for 12 hours, and concanavalin A (ConA) was intravenously injected at 20 mg / kg. After a certain time, blood was collected to prepare serum. After perfusion with PBS, the liver was removed and paraffin blocks and sections were prepared. Alanine transaminase (ALT) in serum was measured using transaminase CII test Wako. Paraffin sections were stained with hematoxylin and eosin and observed using an optical microscope.
野生型Balb/cマウス(♂、9~10週齢)に試験化合物(1.5 mM)を充填した小型浸透圧ポンプ(排出速度1.0μL/hr, 7日間)を腹腔に埋め込むことにより、LPS(18:1)、実施例1の化合物(deoxy-LPS(18:1))または実施例19(LPalloT(18:1))を持続的に供給した。小型浸透圧ポンプ(Alzet社, model 2001)は生理食塩水(大塚生食注)に浸し、37℃で4時間インキュベートしてから用い、それ以外は試験例2と同様の手順で小型浸透圧ポンプをマウスの腹腔に埋め込んだ。4日後に12時間断食し、コンカナバリンA(ConA)を20 mg/kgで尾静注した。一定時間後、採血を行い、血清を調製した。PBSにて灌流後、肝臓を摘出し、パラフィンブロック、切片の作製を行った。トランスアミナーゼCIIテストワコーを用いて血清中のアラニントランスアミナーゼ(ALT)を測定した。パラフィン切片はヘマトキシリンエオジン染色を行い、光学顕微鏡を用いて観察した。 [Test Example 3] Evaluation in a human autoimmune disease hepatitis model mouse A small osmotic pump (discharge rate 1.0 μL / hr) filled with a test compound (1.5 mM) in a wild-type Balb / c mouse (♂, 9 to 10 weeks old) , 7 days) continuously implanted LPS (18: 1), compound of Example 1 (deoxy-LPS (18: 1)) or Example 19 (LPalloT (18: 1)) did. The small osmotic pump (Alzet, model 2001) is immersed in physiological saline (Otsuka raw food injection) and incubated at 37 ° C for 4 hours before use. Otherwise, the small osmotic pump is used in the same procedure as in Test Example 2. Implanted in the abdominal cavity of mice. After 4 days, the animals were fasted for 12 hours, and concanavalin A (ConA) was intravenously injected at 20 mg / kg. After a certain time, blood was collected to prepare serum. After perfusion with PBS, the liver was removed and paraffin blocks and sections were prepared. Alanine transaminase (ALT) in serum was measured using transaminase CII test Wako. Paraffin sections were stained with hematoxylin and eosin and observed using an optical microscope.
試験結果を図9~13に示す。試験結果より、LPS(18:1)、deoxy-LPS(18:1)は血中アラニントランスアミナーゼ(ALT)値の上昇抑制(図9)、肝障害の抑制(図10~14)など、ConA誘発性肝炎の発症を抑制することが明らかとなった。
The test results are shown in FIGS. From the test results, LPS (18: 1) and deoxy-LPS (18: 1) induced ConA, including suppression of blood alanine transaminase (ALT) levels (Fig. 9) and liver damage (Figs. 10-14). It became clear that the onset of sexual hepatitis was suppressed.
[試験例4]活性化リンパ球におけるP2Y10受容体、GPR174受容体の発現量の変化
マウス脾臓細胞をBalb/cマウス(雄、体重20~25g)の脾臓から採取し、RPMI1640培地(100mU/ml ペニシリン、100μg/ml ストレプトマイシン、2mM L-グルタミン、10% FBSを含む)に懸濁させ、24 well plateに1x106個/wellとなるように播種した。リンパ球のマイトジェンである1μg/mlのコンカナバリンA(ConA)存在下で刺激し、37℃で細胞を培養した。一定時間後、ConAで活性化させた脾臓細胞を回収しGenElute Mammalian Total RNA kit (SIGMA-ALDRICH)を用いてtotal RNAを抽出した。また、High capacity cDNA RT kit(Applied Biosystems)を用いて、定法に従って逆転写させ、cDNAを得た。cDNAと、目的遺伝子を計測するためのプライマーおよびSYBR Green PCR Master Mix(Applied Biosystems)を混合し、7300 Real-Time PCR Systemを用いてリアルタイムPCR法によりPCR産物を定量した。プライマーとして配列番号25~28に示すオリゴヌクレオチドを使用した。またコントロールジーンとして使用したGAPDHのプライマーを配列番号29および30に示す。P2Y10受容体、GPR174受容体の発現量について図15および16に示す。何れの受容体もコンカナバリンAによる刺激で発現量が増加することが確認された。 [Test Example 4] Changes in expression levels of P2Y10 receptor and GPR174 receptor in activated lymphocytes Mouse spleen cells were collected from the spleen of Balb / c mice (male, body weight 20-25 g) and RPMI1640 medium (100 mU / ml). Penicillin, 100 μg / ml streptomycin, 2 mM L-glutamine, 10% FBS), and seeded on a 24-well plate at 1 × 10 6 cells / well. Cells were stimulated in the presence of 1 μg / ml concanavalin A (ConA), a lymphocyte mitogen, and cultured at 37 ° C. After a certain time, spleen cells activated with ConA were collected, and total RNA was extracted using GenElute Mammalian Total RNA kit (SIGMA-ALDRICH). Further, cDNA was obtained by reverse transcription using a high capacity cDNA RT kit (Applied Biosystems) according to a conventional method. cDNA, a primer for measuring a target gene, and SYBR Green PCR Master Mix (Applied Biosystems) were mixed, and PCR products were quantified by real-time PCR using 7300 Real-Time PCR System. Oligonucleotides shown in SEQ ID NOs: 25 to 28 were used as primers. Further, GAPDH primers used as control genes are shown in SEQ ID NOs: 29 and 30. The expression levels of P2Y10 receptor and GPR174 receptor are shown in FIGS. It was confirmed that the expression level of any receptor increased upon stimulation with concanavalin A.
マウス脾臓細胞をBalb/cマウス(雄、体重20~25g)の脾臓から採取し、RPMI1640培地(100mU/ml ペニシリン、100μg/ml ストレプトマイシン、2mM L-グルタミン、10% FBSを含む)に懸濁させ、24 well plateに1x106個/wellとなるように播種した。リンパ球のマイトジェンである1μg/mlのコンカナバリンA(ConA)存在下で刺激し、37℃で細胞を培養した。一定時間後、ConAで活性化させた脾臓細胞を回収しGenElute Mammalian Total RNA kit (SIGMA-ALDRICH)を用いてtotal RNAを抽出した。また、High capacity cDNA RT kit(Applied Biosystems)を用いて、定法に従って逆転写させ、cDNAを得た。cDNAと、目的遺伝子を計測するためのプライマーおよびSYBR Green PCR Master Mix(Applied Biosystems)を混合し、7300 Real-Time PCR Systemを用いてリアルタイムPCR法によりPCR産物を定量した。プライマーとして配列番号25~28に示すオリゴヌクレオチドを使用した。またコントロールジーンとして使用したGAPDHのプライマーを配列番号29および30に示す。P2Y10受容体、GPR174受容体の発現量について図15および16に示す。何れの受容体もコンカナバリンAによる刺激で発現量が増加することが確認された。 [Test Example 4] Changes in expression levels of P2Y10 receptor and GPR174 receptor in activated lymphocytes Mouse spleen cells were collected from the spleen of Balb / c mice (male, body weight 20-25 g) and RPMI1640 medium (100 mU / ml). Penicillin, 100 μg / ml streptomycin, 2 mM L-glutamine, 10% FBS), and seeded on a 24-well plate at 1 × 10 6 cells / well. Cells were stimulated in the presence of 1 μg / ml concanavalin A (ConA), a lymphocyte mitogen, and cultured at 37 ° C. After a certain time, spleen cells activated with ConA were collected, and total RNA was extracted using GenElute Mammalian Total RNA kit (SIGMA-ALDRICH). Further, cDNA was obtained by reverse transcription using a high capacity cDNA RT kit (Applied Biosystems) according to a conventional method. cDNA, a primer for measuring a target gene, and SYBR Green PCR Master Mix (Applied Biosystems) were mixed, and PCR products were quantified by real-time PCR using 7300 Real-Time PCR System. Oligonucleotides shown in SEQ ID NOs: 25 to 28 were used as primers. Further, GAPDH primers used as control genes are shown in SEQ ID NOs: 29 and 30. The expression levels of P2Y10 receptor and GPR174 receptor are shown in FIGS. It was confirmed that the expression level of any receptor increased upon stimulation with concanavalin A.
[試験例5]マウス脾臓細胞のICAM-1に対する接着能の評価
マウス脾臓細胞をBalb/cマウス(雄、体重20~25 g)の脾臓から採取し、RPMI1640培地(100 mU/ml ペニシリン、100μg/ml ストレプトマイシン、2 mM L-グルタミン、10% FBSを含む)に懸濁させ、24 well plateに5 x 106個/wellとなるように播種した。リンパ球のマイトジェンであるコンカナバリンA(ConA、1μg/ml)存在下で刺激し、37℃で48時間細胞を培養した。ConAで活性化させた脾臓細胞を回収し、PBSで2回洗浄し、1μMのCMFDAを含むPBSに懸濁させ、37℃で1時間蛍光標識した。さらに、脾臓細胞をPBSで2回洗浄し、RPMI1640培地(0.01%のウシ血清アルブミン(BSA)を含む)に懸濁させた。2.5μg/mlのリコンビナントマウスICAM Fcで96 well black plateを4℃で24時間処理し、0.5% BSAを含むPBSで30分間ブロッキングを行った。96 well palteをPBSで3回洗浄し、脾臓細胞を2 x 105個/wellとなるように播種した。試験化合物の存在下、蛍光標識した脾臓細胞を37℃で、リンパ球の接着を促進することが知られている10 mMのMg2+と1 mMのEGTAで刺激した。30分後、96 well plateを逆さにして30分間静置させ、接着していない細胞を除去し、1% NP-40を含むPBSを50μlずつ加えて細胞を溶解させた。プレートに接着した細胞の数は蛍光プレートリーダーを用いて蛍光強度を測定することで決定された。接着した細胞の割合を全細胞数のパーセンテージとして計算した。測定値はn=3で行った実験の平均値±標準誤差として示した。 [Test Example 5] Evaluation of adhesion ability of mouse spleen cells to ICAM-1 Mouse spleen cells were collected from the spleen of Balb / c mice (male, body weight 20-25 g) and RPMI1640 medium (100 mU / ml penicillin, 100 μg). / ml streptomycin, 2 mM L-glutamine, 10% FBS) and seeded on a 24 well plate at 5 × 10 6 cells / well. Stimulation was performed in the presence of concanavalin A (ConA, 1 μg / ml), a lymphocyte mitogen, and the cells were cultured at 37 ° C. for 48 hours. Spleen cells activated with ConA were collected, washed twice with PBS, suspended in PBS containing 1 μM CMFDA, and fluorescently labeled at 37 ° C. for 1 hour. Further, the spleen cells were washed twice with PBS and suspended in RPMI1640 medium (containing 0.01% bovine serum albumin (BSA)). A 96-well black plate was treated with 2.5 μg / ml recombinant mouse ICAM Fc at 4 ° C. for 24 hours, and blocked with PBS containing 0.5% BSA for 30 minutes. 96 well palte was washed 3 times with PBS, and spleen cells were seeded at 2 × 10 5 cells / well. In the presence of test compounds, fluorescently labeled spleen cells were stimulated at 37 ° C. with 10 mM Mg 2+ and 1 mM EGTA, which are known to promote lymphocyte adhesion. After 30 minutes, the 96-well plate was inverted and allowed to stand for 30 minutes to remove non-adherent cells, and 50 μl of PBS containing 1% NP-40 was added to lyse the cells. The number of cells attached to the plate was determined by measuring the fluorescence intensity using a fluorescence plate reader. The percentage of cells that adhered was calculated as a percentage of the total number of cells. The measured value is shown as an average value ± standard error of an experiment conducted at n = 3.
マウス脾臓細胞をBalb/cマウス(雄、体重20~25 g)の脾臓から採取し、RPMI1640培地(100 mU/ml ペニシリン、100μg/ml ストレプトマイシン、2 mM L-グルタミン、10% FBSを含む)に懸濁させ、24 well plateに5 x 106個/wellとなるように播種した。リンパ球のマイトジェンであるコンカナバリンA(ConA、1μg/ml)存在下で刺激し、37℃で48時間細胞を培養した。ConAで活性化させた脾臓細胞を回収し、PBSで2回洗浄し、1μMのCMFDAを含むPBSに懸濁させ、37℃で1時間蛍光標識した。さらに、脾臓細胞をPBSで2回洗浄し、RPMI1640培地(0.01%のウシ血清アルブミン(BSA)を含む)に懸濁させた。2.5μg/mlのリコンビナントマウスICAM Fcで96 well black plateを4℃で24時間処理し、0.5% BSAを含むPBSで30分間ブロッキングを行った。96 well palteをPBSで3回洗浄し、脾臓細胞を2 x 105個/wellとなるように播種した。試験化合物の存在下、蛍光標識した脾臓細胞を37℃で、リンパ球の接着を促進することが知られている10 mMのMg2+と1 mMのEGTAで刺激した。30分後、96 well plateを逆さにして30分間静置させ、接着していない細胞を除去し、1% NP-40を含むPBSを50μlずつ加えて細胞を溶解させた。プレートに接着した細胞の数は蛍光プレートリーダーを用いて蛍光強度を測定することで決定された。接着した細胞の割合を全細胞数のパーセンテージとして計算した。測定値はn=3で行った実験の平均値±標準誤差として示した。 [Test Example 5] Evaluation of adhesion ability of mouse spleen cells to ICAM-1 Mouse spleen cells were collected from the spleen of Balb / c mice (male, body weight 20-25 g) and RPMI1640 medium (100 mU / ml penicillin, 100 μg). / ml streptomycin, 2 mM L-glutamine, 10% FBS) and seeded on a 24 well plate at 5 × 10 6 cells / well. Stimulation was performed in the presence of concanavalin A (ConA, 1 μg / ml), a lymphocyte mitogen, and the cells were cultured at 37 ° C. for 48 hours. Spleen cells activated with ConA were collected, washed twice with PBS, suspended in PBS containing 1 μM CMFDA, and fluorescently labeled at 37 ° C. for 1 hour. Further, the spleen cells were washed twice with PBS and suspended in RPMI1640 medium (containing 0.01% bovine serum albumin (BSA)). A 96-well black plate was treated with 2.5 μg / ml recombinant mouse ICAM Fc at 4 ° C. for 24 hours, and blocked with PBS containing 0.5% BSA for 30 minutes. 96 well palte was washed 3 times with PBS, and spleen cells were seeded at 2 × 10 5 cells / well. In the presence of test compounds, fluorescently labeled spleen cells were stimulated at 37 ° C. with 10 mM Mg 2+ and 1 mM EGTA, which are known to promote lymphocyte adhesion. After 30 minutes, the 96-well plate was inverted and allowed to stand for 30 minutes to remove non-adherent cells, and 50 μl of PBS containing 1% NP-40 was added to lyse the cells. The number of cells attached to the plate was determined by measuring the fluorescence intensity using a fluorescence plate reader. The percentage of cells that adhered was calculated as a percentage of the total number of cells. The measured value is shown as an average value ± standard error of an experiment conducted at n = 3.
上記実施例で調製した化合物のうち、LPS(18:1)、deoxy LPS(18:1)、LPalloT(18:1)を使用した。試験結果を図17に示す。試験結果より、LPS(18:1)、deoxy LPS(18:1)は脾臓細胞のICAM-1に対する接着を抑制するが、LPalloT(18:1)はこの接着を抑制しないことが確認された(化合物終濃度:10μM)。
Among the compounds prepared in the above examples, LPS (18: 1), deoxy LPS (18: 1), and LPalloT (18: 1) were used. The test results are shown in FIG. From the test results, it was confirmed that LPS (18: 1) and deoxy LPS (18: 1) inhibit adhesion of spleen cells to ICAM-1, but LPalloT (18: 1) does not inhibit this adhesion ( Compound final concentration: 10 μM).
[試験例6]マウス脾臓細胞のIL-2産生に対する抑制効果の評価
マウス脾臓細胞をBalb/cマウス(雄、体重20~25g)の脾臓から採取し、RPMI1640培地(100 mU/ml ペニシリン、100μg/ml ストレプトマイシン、2 mM L-グルタミン、10% FBSを含む)に懸濁させ、24 well plateに1 x 106個/wellとなるように播種した。1μg/mlのConA存在下で刺激し、37 ℃で60時間細胞を培養した。ConAで活性化させた脾臓細胞を回収し、RPMI1640培地(0.01%のBSAを含む)に懸濁させ、96 well plateに5 x 105個/wellとなるように播種した。試験化合物の存在下、活性化させた脾臓細胞を37℃で、IL-2の産生を促進させるために1μ/mlのConAで刺激した。12時間後、96 well plateから培養上清を回収した。上清中のIL-2の濃度は、Mouse IL-2 Matched Antibody Pairs for ELISA(eBioscience,BMS601MST)を用いて、サンドイッチELISAにより決定された。測定値は、異なるマウス個体から採取した脾臓細胞を用いて数回の反復実験を行い、平均値±標準誤差として示した。 [Test Example 6] Evaluation of inhibitory effect of mouse spleen cells on IL-2 production Mouse spleen cells were collected from the spleen of Balb / c mice (male, body weight 20-25 g), and RPMI1640 medium (100 mU / ml penicillin, 100 μg). / ml streptomycin, 2 mM L-glutamine, 10% FBS), and seeded on a 24-well plate at 1 × 10 6 cells / well. Stimulation was performed in the presence of 1 μg / ml ConA, and the cells were cultured at 37 ° C. for 60 hours. Spleen cells activated with ConA were collected, suspended in RPMI1640 medium (containing 0.01% BSA), and seeded on a 96-well plate at 5 × 10 5 cells / well. Activated spleen cells in the presence of test compounds were stimulated at 37 ° C. with 1 μ / ml ConA to promote IL-2 production. After 12 hours, the culture supernatant was collected from the 96 well plate. The concentration of IL-2 in the supernatant was determined by sandwich ELISA using Mouse IL-2 Matched Antibody Pairs for ELISA (eBioscience, BMS601MST). The measured value was expressed as mean value ± standard error after several repeated experiments using spleen cells collected from different mouse individuals.
マウス脾臓細胞をBalb/cマウス(雄、体重20~25g)の脾臓から採取し、RPMI1640培地(100 mU/ml ペニシリン、100μg/ml ストレプトマイシン、2 mM L-グルタミン、10% FBSを含む)に懸濁させ、24 well plateに1 x 106個/wellとなるように播種した。1μg/mlのConA存在下で刺激し、37 ℃で60時間細胞を培養した。ConAで活性化させた脾臓細胞を回収し、RPMI1640培地(0.01%のBSAを含む)に懸濁させ、96 well plateに5 x 105個/wellとなるように播種した。試験化合物の存在下、活性化させた脾臓細胞を37℃で、IL-2の産生を促進させるために1μ/mlのConAで刺激した。12時間後、96 well plateから培養上清を回収した。上清中のIL-2の濃度は、Mouse IL-2 Matched Antibody Pairs for ELISA(eBioscience,BMS601MST)を用いて、サンドイッチELISAにより決定された。測定値は、異なるマウス個体から採取した脾臓細胞を用いて数回の反復実験を行い、平均値±標準誤差として示した。 [Test Example 6] Evaluation of inhibitory effect of mouse spleen cells on IL-2 production Mouse spleen cells were collected from the spleen of Balb / c mice (male, body weight 20-25 g), and RPMI1640 medium (100 mU / ml penicillin, 100 μg). / ml streptomycin, 2 mM L-glutamine, 10% FBS), and seeded on a 24-well plate at 1 × 10 6 cells / well. Stimulation was performed in the presence of 1 μg / ml ConA, and the cells were cultured at 37 ° C. for 60 hours. Spleen cells activated with ConA were collected, suspended in RPMI1640 medium (containing 0.01% BSA), and seeded on a 96-well plate at 5 × 10 5 cells / well. Activated spleen cells in the presence of test compounds were stimulated at 37 ° C. with 1 μ / ml ConA to promote IL-2 production. After 12 hours, the culture supernatant was collected from the 96 well plate. The concentration of IL-2 in the supernatant was determined by sandwich ELISA using Mouse IL-2 Matched Antibody Pairs for ELISA (eBioscience, BMS601MST). The measured value was expressed as mean value ± standard error after several repeated experiments using spleen cells collected from different mouse individuals.
上記実施例で調製した化合物のうち、LPS(18:1)、deoxy-LPS(18:1)、LPalloT(18:1)を使用した。試験結果を図18に示す。試験結果より、LPS(18:1)、LPalloT(18:1)は濃度依存的に脾臓細胞のIL-2産生を抑制するが、deoxy-LPS(18:1)はIL-2の産生を抑制しないことが確認された。
Among the compounds prepared in the above examples, LPS (18: 1), deoxy-LPS (18: 1), and LPalloT (18: 1) were used. The test results are shown in FIG. From the test results, LPS (18: 1) and LPalloT (18: 1) inhibit IL-2 production in spleen cells in a concentration-dependent manner, but deoxy-LPS (18: 1) inhibits IL-2 production. It was confirmed not to.
Claims (20)
- 式(I):
R1は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、またはベンジルであり;
R2およびR3は、それぞれ独立に、水素原子、C1-6アルキル、ホルミル、C1-6アルキルカルボニル、C1-6アルコキシカルボニル、C1-6アルコキシC1-6アルキル、またはベンジルオキシカルボニルから選択され;
R4は、水素原子またはC1-3アルキルであり;
R5は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、C1-6アルキルカルボニルオキシC1-6アルキル、C1-6アルコキシカルボニルオキシC1-6アルキル、フェニル、ベンジル、フタリジル、ジオキソレノンイルメチル、またはフリルメチルであり;
Xは、水素原子、ハロゲン原子、ヒドロキシ、C1-6アルコキシ、ホルミルオキシ、C1-6アルキルカルボニルオキシ、またはベンジルオキシであり;
nは、0~4から選択される整数であり;
Yは、-O-、-C(=O)O-、-OC(=O)-、-C(=O)NR7-、または-NR7C(=O)-であり;
R6は、C3-30アルキルであり、ここでアルキルには1以上のフェニレンまたは5-6員ヘテロアリーレンが挿入されていてもよく、アルキルの1以上の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1以上の単結合は、二重結合または三重結合に置き換えられていてもよく、アルキルの末端はフェニルまたは5-6員ヘテロアリールにより置換されていてもよく、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシおよびハロゲン原子から選択される1以上の置換基により置換されていてもよく;
R7は、水素原子、またはC1-6アルキルであり;
上記のベンジル、ベンジルオキシ、フェニル、フタリジル、ジオキソレノンイルメチル、フリルメチル、フェニレン、およびヘテロアリーレンは、C1-6アルキル、C1-6アルコキシ、ヒドロキシ、C1-6アルキルカルボニルオキシ、ニトロ、フェニルおよびハロゲン原子から選択される1以上の置換基により置換されていてもよい]
で表される化合物、または医薬として許容なその塩を含有する、自己免疫疾患の治療剤または予防剤。 Formula (I):
R 1 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, or benzyl;
R 2 and R 3 are each independently a hydrogen atom, C 1-6 alkyl, formyl, C 1-6 alkylcarbonyl, C 1-6 alkoxycarbonyl, C 1-6 alkoxy C 1-6 alkyl, or benzyloxy Selected from carbonyl;
R 4 is a hydrogen atom or C 1-3 alkyl;
R 5 represents a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, C 1-6 alkylcarbonyloxy C 1-6 alkyl, C 1-6 alkoxycarbonyloxy C 1-6 alkyl, Phenyl, benzyl, phthalidyl, dioxolenoneylmethyl, or furylmethyl;
X is a hydrogen atom, a halogen atom, hydroxy, C 1-6 alkoxy, formyloxy, C 1-6 alkylcarbonyloxy, or benzyloxy;
n is an integer selected from 0 to 4;
Y is —O—, —C (═O) O—, —OC (═O) —, —C (═O) NR 7 —, or —NR 7 C (═O) —;
R 6 is C 3-30 alkyl, wherein one or more phenylene or 5-6 membered heteroarylene may be inserted in the alkyl, and one or more —CH 2 — of alkyl is —O— and One or more single bonds between carbon atoms may be replaced by double bonds or triple bonds, and the alkyl ends may be substituted by phenyl or 5-6 membered heteroaryl. Well, the phenyl or heteroaryl may be substituted with one or more substituents selected from C 1-10 alkyl, C 1-10 alkoxy and a halogen atom;
R 7 is a hydrogen atom or C 1-6 alkyl;
The above benzyl, benzyloxy, phenyl, phthalidyl, dioxolenoneylmethyl, furylmethyl, phenylene, and heteroarylene are C 1-6 alkyl, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro May be substituted with one or more substituents selected from phenyl and halogen atoms]
The therapeutic agent or preventive agent of an autoimmune disease containing the compound represented by these, or its pharmaceutically acceptable salt. - R6が、C10-24アルキル(ここでアルキルの1以上の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1以上の単結合は、二重結合または三重結合に置き換えられていてもよい)、または式:-(CH2)m-Qから選択され;
mは、2~20から選択される整数であり;
Qは、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-10アルキル、フェニルC1-10アルコキシ、5-6員ヘテロアリールC1-10アルキル、または5-6員ヘテロアリールC1-10アルコキシにより置換されていてもよい、請求項1または2に記載の治療剤または予防剤。 R 6 is C 10-24 alkyl (wherein one or more —CH 2 — of alkyl may be replaced by —O—, and one or more single bonds between carbon atoms are double bonds or triple bonds) Or may be selected from the formula: — (CH 2 ) m —Q;
m is an integer selected from 2 to 20;
Q is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-10 alkyl, phenyl C 1-10 alkoxy, 5-6 The therapeutic or prophylactic agent according to claim 1 or 2, which may be substituted by a member heteroaryl C 1-10 alkyl or a 5-6 membered heteroaryl C 1-10 alkoxy. - R6が、C12-20アルキル(ここでアルキルの1~3の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1つの単結合は、二重結合に置き換えられていてもよい)、または式:-(CH2)m-Qから選択され;
mは、2~7から選択される整数であり;
Qは、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-3アルキル、フェニルC1-3アルコキシ、5-6員ヘテロアリールC1-3アルキル、または5-6員ヘテロアリールC1-3アルコキシにより置換されていてもよい、請求項3に記載の治療剤または予防剤。 R 6 is C 12-20 alkyl (wherein 1-3 of alkyl —CH 2 — may be replaced by —O—, and one single bond between carbon atoms is replaced by a double bond) Or selected from the formula: — (CH 2 ) m —Q;
m is an integer selected from 2 to 7;
Q is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5-6 The therapeutic or prophylactic agent according to claim 3, which may be substituted by a member heteroaryl C 1-3 alkyl or a 5-6 membered heteroaryl C 1-3 alkoxy. - (2S)-2-アミノ-3-[(3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-バセノイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-エライドイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-パルミトレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-ミリストイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-ラウロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-ヘキサデシルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(2-オレイルオキシエトキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(3-オレイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(4-オレイルオキシブトキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[(5-オレイルオキシペントキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[3-{8-(4-ヘプチル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[3-{8-(4-ヘキシル-1,2,3-トリアゾル-1-イル)オクタノイルオキシ}プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[6-{4-ヘキシルオキシ-(2Z)-2-ブテン-1-イルオキシ}ヘキサノイルオキシ]プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[2-(オレイルオキシカルボニル)エトキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[3-(オレイルオキシカルボニル)プロポキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[4-(オレイルオキシカルボニル)ブトキシ-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S,3S)-2-アミノ-3-[(3-オレオイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[{3-((9E)-オクタデセノイルオキシ)プロピル}-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[(3-ステアロイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[(3-パルミトレオイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[(3-パルミトイルオキシプロピル)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸;
(2S,3S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]酪酸;
(2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2S)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(2-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(3-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(4-ヘプチルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-{3-(4-ベンジルオキシフェニル)プロピオニルオキシ}プロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-オレオイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-ステアロイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸;および
(2S)-2-アミノ-3-[((2R)-2-ヒドロキシ-3-パルミトイルオキシプロポキシ)-ヒドロキシホスホリルオキシ]プロピオン酸
から選択される化合物、または医薬として許容なその塩を含有する、請求項1~4のいずれか1項に記載の治療剤または予防剤。 (2S) -2-amino-3-[(3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3-[(3-basenoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-Elideyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-palmitoleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-myristoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-lauroyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-hexadecyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(2-oleyloxyethoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(3-oleyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(4-oleyloxybutoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[(5-oleyloxypentoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [3- {8- (4-heptyl-1,2,3-triazol-1-yl) octanoyloxy} propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [3- {8- (4-hexyl-1,2,3-triazol-1-yl) octanoyloxy} propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [6- {4-hexyloxy- (2Z) -2-buten-1-yloxy} hexanoyloxy] propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [2- (oleyloxycarbonyl) ethoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [3- (oleyloxycarbonyl) propoxy-hydroxyphosphoryloxy] propionic acid;
(2S) -2-Amino-3- [4- (oleyloxycarbonyl) butoxy-hydroxyphosphoryloxy] propionic acid;
(2S, 3S) -2-Amino-3-[(3-oleoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[{3-((9E) -octadecenoyloxy) propyl} -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-Amino-3-[(3-stearoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[(3-palmitoleoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-Amino-3-[(3-palmitoyloxypropyl) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid;
(2S, 3S) -2-amino-3-[((2R) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] butyric acid;
(2S) -2-amino-3-[((2S) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2S) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2S) -2-hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (2-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (3-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (4-heptyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3- {3- (4-benzyloxyphenyl) propionyloxy} propoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3-oleoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid;
(2S) -2-amino-3-[((2R) -2-hydroxy-3-stearoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid; and (2S) -2-amino-3-[((2R) The therapeutic agent according to any one of claims 1 to 4, comprising a compound selected from -2-hydroxy-3-palmitoyloxypropoxy) -hydroxyphosphoryloxy] propionic acid, or a pharmaceutically acceptable salt thereof. Or prophylactic agent. - 自己免疫疾患が、悪性関節リウマチ、全身エリテマトーデス、多発性硬化症、重症筋無力症、バセドウ病、抗リン脂質抗体症候群、シェーグレン症候群、原発性胆汁性肝硬変、多発性筋炎、および自己免疫性肝炎から選択される疾患である、請求項1~5のいずれか1項に記載の治療剤または予防剤。 Autoimmune diseases from malignant rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, Graves' disease, antiphospholipid syndrome, Sjogren's syndrome, primary biliary cirrhosis, polymyositis, and autoimmune hepatitis The therapeutic or prophylactic agent according to any one of claims 1 to 5, which is a selected disease.
- 式(Ia):
R11は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、またはベンジルであり;
R12およびR13は、それぞれ独立に、水素原子、C1-6アルキル、ホルミル、C1-6アルキルカルボニル、C1-6アルコキシカルボニル、C1-6アルコキシC1-6アルキル、またはベンジルオキシカルボニルから選択され;
R15は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、C1-6アルキルカルボニルオキシC1-6アルキル、C1-6アルコキシカルボニルオキシC1-6アルキル、フェニル、ベンジル、フタリジル、ジオキソレノンイルメチル、またはフリルメチルであり;
pは、2~6から選択される整数であり;
Y1は、直接結合、または-C(=O)-であり;
R16は、C3-30アルキルであり、ここでアルキルには1以上のフェニレンまたは5-6員ヘテロアリーレンが挿入されていてもよく、アルキルの1以上の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1以上の単結合は、二重結合または三重結合に置き換えられていてもよく、アルキルの末端はフェニルまたは5-6員ヘテロアリールにより置換されていてもよく、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシおよびハロゲン原子から選択される1以上の置換基により置換されていてもよく;
上記のベンジル、ベンジルオキシ、フェニル、フタリジル、ジオキソレノンイルメチル、フリルメチル、フェニレン、およびヘテロアリーレンは、C1-6アルキル、C1-6アルコキシ、ヒドロキシ、C1-6アルキルカルボニルオキシ、ニトロ、フェニルおよびハロゲン原子から選択される1以上の置換基により置換されていてもよい]
で表される化合物、または医薬として許容なその塩。 Formula (Ia):
R 11 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, or benzyl;
R 12 and R 13 are each independently a hydrogen atom, C 1-6 alkyl, formyl, C 1-6 alkylcarbonyl, C 1-6 alkoxycarbonyl, C 1-6 alkoxy C 1-6 alkyl, or benzyloxy Selected from carbonyl;
R 15 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, C 1-6 alkylcarbonyloxy C 1-6 alkyl, C 1-6 alkoxycarbonyloxy C 1-6 alkyl, Phenyl, benzyl, phthalidyl, dioxolenoneylmethyl, or furylmethyl;
p is an integer selected from 2 to 6;
Y 1 is a direct bond or —C (═O) —;
R 16 is C 3-30 alkyl, wherein one or more phenylene or 5-6 membered heteroarylene may be inserted in the alkyl, and one or more —CH 2 — of alkyl is —O— and One or more single bonds between carbon atoms may be replaced by double bonds or triple bonds, and the alkyl ends may be substituted by phenyl or 5-6 membered heteroaryl. Well, the phenyl or heteroaryl may be substituted with one or more substituents selected from C 1-10 alkyl, C 1-10 alkoxy and a halogen atom;
The above benzyl, benzyloxy, phenyl, phthalidyl, dioxolenoneylmethyl, furylmethyl, phenylene, and heteroarylene are C 1-6 alkyl, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro May be substituted with one or more substituents selected from phenyl and halogen atoms]
Or a pharmaceutically acceptable salt thereof. - R16が、C10-24アルキル(ここでアルキルの1以上の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1以上の単結合は、二重結合または三重結合に置き換えられていてもよい)、または式:-(CH2)r-Q1から選択され;
rは、2~20から選択される整数であり;
Q1は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-10アルキル、フェニルC1-10アルコキシ、5-6員ヘテロアリールC1-10アルキル、または5-6員ヘテロアリールC1-10アルコキシにより置換されていてもよい、請求項7に記載の化合物、または医薬として許容なその塩。 R 16 is C 10-24 alkyl (wherein one or more —CH 2 — of alkyl may be replaced by —O—, and one or more single bonds between carbon atoms are double bonds or triple bonds) Or is selected from the formula: — (CH 2 ) r —Q 1 ;
r is an integer selected from 2 to 20;
Q 1 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-10 alkyl, phenyl C 1-10 alkoxy, 5- 8. The compound according to claim 7, or a pharmaceutically acceptable salt thereof, optionally substituted by 6-membered heteroaryl C 1-10 alkyl, or 5-6-membered heteroaryl C 1-10 alkoxy. - R16が、C12-20アルキル(ここでアルキルの1~3の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1つの単結合は、二重結合に置き換えられていてもよい)、または式:-(CH2)r-Q1から選択され;
rは、2~7から選択される整数であり;
Q1は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-3アルキル、フェニルC1-3アルコキシ、5-6員ヘテロアリールC1-3アルキル、または5-6員ヘテロアリールC1-3アルコキシにより置換されていてもよい、請求項8に記載の化合物、または医薬として許容なその塩。 R 16 is C 12-20 alkyl (wherein 1 to 3 —CH 2 — of alkyl may be replaced by —O—, and one single bond between carbon atoms is replaced by a double bond. Or selected from the formula: — (CH 2 ) r —Q 1 ;
r is an integer selected from 2 to 7;
Q 1 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5- 9. The compound according to claim 8, or a pharmaceutically acceptable salt thereof, optionally substituted by 6-membered heteroaryl C 1-3 alkyl, or 5-6-membered heteroaryl C 1-3 alkoxy. - 式(Ib):
R21は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、またはベンジルであり;
R22およびR23は、それぞれ独立に、水素原子、C1-6アルキル、ホルミル、C1-6アルキルカルボニル、C1-6アルコキシカルボニル、C1-6アルコキシC1-6アルキル、またはベンジルオキシカルボニルから選択され;
R24は、水素原子またはC1-3アルキルであり;
R25は、水素原子、C1-6アルキル、C1-6アルコキシC1-6アルキル、C1-6アルキルカルボニルオキシC1-6アルキル、C1-6アルコキシカルボニルオキシC1-6アルキル、フェニル、ベンジル、フタリジル、ジオキソレノンイルメチル、またはフリルメチルであり;
X2は、水素原子、ハロゲン原子、ヒドロキシ、C1-6アルコキシ、ホルミルオキシ、C1-6アルキルカルボニルオキシ、またはベンジルオキシであり;
sは、0~4から選択される整数であり;
Y2は、-O-、-C(=O)O-、-OC(=O)-、-C(=O)NR27-、または-NR27C(=O)-であり;
R26は、式:-(CH2)t-Q2であり;
tは、2~20から選択される整数であり;
Q2は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-10アルキル、フェニルC1-10アルコキシ、5-6員ヘテロアリールC1-10アルキル、または5-6員ヘテロアリールC1-10アルコキシにより置換されていてもよく;
R27は、水素原子、またはC1-6アルキルであり;
さらに上記のベンジル、ベンジルオキシ、フェニル、フタリジル、ジオキソレノンイルメチル、およびフリルメチルは、C1-6アルキル、C1-6アルコキシ、ヒドロキシ、C1-6アルキルカルボニルオキシ、ニトロ、フェニルおよびハロゲン原子から選択される1以上の置換基により置換されていてもよい]
で表される化合物、または医薬として許容なその塩。 Formula (Ib):
R 21 is a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, or benzyl;
R 22 and R 23 are each independently a hydrogen atom, C 1-6 alkyl, formyl, C 1-6 alkylcarbonyl, C 1-6 alkoxycarbonyl, C 1-6 alkoxy C 1-6 alkyl, or benzyloxy Selected from carbonyl;
R 24 is a hydrogen atom or C 1-3 alkyl;
R 25 represents a hydrogen atom, C 1-6 alkyl, C 1-6 alkoxy C 1-6 alkyl, C 1-6 alkylcarbonyloxy C 1-6 alkyl, C 1-6 alkoxycarbonyloxy C 1-6 alkyl, Phenyl, benzyl, phthalidyl, dioxolenoneylmethyl, or furylmethyl;
X 2 is a hydrogen atom, a halogen atom, hydroxy, C 1-6 alkoxy, formyloxy, C 1-6 alkylcarbonyloxy, or benzyloxy;
s is an integer selected from 0 to 4;
Y 2 is —O—, —C (═O) O—, —OC (═O) —, —C (═O) NR 27 —, or —NR 27 C (═O) —;
R 26 is of the formula: — (CH 2 ) t —Q 2 ;
t is an integer selected from 2 to 20;
Q 2 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-10 alkyl, phenyl C 1-10 alkoxy, 5- Optionally substituted by 6-membered heteroaryl C 1-10 alkyl, or 5-6-membered heteroaryl C 1-10 alkoxy;
R 27 is a hydrogen atom or C 1-6 alkyl;
Further, the above benzyl, benzyloxy, phenyl, phthalidyl, dioxolenoneylmethyl and furylmethyl are C 1-6 alkyl, C 1-6 alkoxy, hydroxy, C 1-6 alkylcarbonyloxy, nitro, phenyl and halogen. Optionally substituted by one or more substituents selected from atoms]
Or a pharmaceutically acceptable salt thereof. - R26が、C12-20アルキル(ここでアルキルの1~3の-CH2-は-O-と置き換えられていてもよく、炭素原子間の1つの単結合は、二重結合に置き換えられていてもよい)、または式:-(CH2)s-Q2から選択され;
sは、2~7から選択される整数であり;
Q2は、フェニルまたは5-6員ヘテロアリールであり、該フェニルまたはヘテロアリールは、C1-10アルキル、C1-10アルコキシ、フェニルC1-3アルキル、フェニルC1-3アルコキシ、5-6員ヘテロアリールC1-3アルキル、または5-6員ヘテロアリールC1-3アルコキシにより置換されていてもよい、請求項10または11に記載の化合物、または医薬として許容なその塩。 R 26 is C 12-20 alkyl (wherein 1 to 3 —CH 2 — in the alkyl may be replaced by —O—, and one single bond between carbon atoms is replaced by a double bond. Or selected from the formula: — (CH 2 ) s —Q 2 ;
s is an integer selected from 2 to 7;
Q 2 is phenyl or 5-6 membered heteroaryl, which is C 1-10 alkyl, C 1-10 alkoxy, phenyl C 1-3 alkyl, phenyl C 1-3 alkoxy, 5- 12. The compound according to claim 10 or 11, or a pharmaceutically acceptable salt thereof, optionally substituted by 6-membered heteroaryl C 1-3 alkyl, or 5-6-membered heteroaryl C 1-3 alkoxy. - P2Y10またはGPR174を使用して、リゾホスファチジルセリン受容体への被検化合物のアゴニスト活性またはアンタゴニスト活性を評価することを含む、自己免疫疾患の治療剤または予防剤のスクリーニング方法。 A screening method for a therapeutic or prophylactic agent for autoimmune diseases, comprising evaluating the agonistic or antagonistic activity of a test compound for lysophosphatidylserine receptors using P2Y10 or GPR174.
- アゴニスト活性を評価する、請求項13に記載の方法。 The method according to claim 13, wherein the agonist activity is evaluated.
- インターロイキン2産生抑制剤をスクリーニングするための、請求項13または14に記載の方法。 The method according to claim 13 or 14, for screening for an interleukin 2 production inhibitor.
- リンパ球接着抑制剤をスクリーニングするための、請求項13または14に記載の方法。 The method according to claim 13 or 14, for screening a lymphocyte adhesion inhibitor.
- P2Y10またはGPR174をコードする遺伝子を発現する細胞を使用する、請求項13~16のいずれか1項に記載の方法。 The method according to any one of claims 13 to 16, wherein a cell expressing a gene encoding P2Y10 or GPR174 is used.
- アルカリフォスファターゼ(AP)で標識化されたトランスフォーミング増殖因子α(TGFα)をコードする遺伝子をさらに発現する細胞を使用する、請求項17に記載の方法。 The method according to claim 17, wherein cells further expressing a gene encoding transforming growth factor α (TGFα) labeled with alkaline phosphatase (AP) are used.
- 対照化合物としてリゾホスファチジルセリンまたはその類縁体を使用する、請求項13~18のいずれか1項に記載の方法。 The method according to any one of claims 13 to 18, wherein lysophosphatidylserine or an analog thereof is used as a control compound.
- 自己免疫疾患が、悪性関節リウマチ、全身エリテマトーデス、多発性硬化症、重症筋無力症、バセドウ病、抗リン脂質抗体症候群、シェーグレン症候群、原発性胆汁性肝硬変、多発性筋炎、および自己免疫性肝炎から選択される疾患である、請求項13~19のいずれか1項に記載の方法。 Autoimmune diseases from malignant rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, Graves' disease, antiphospholipid syndrome, Sjogren's syndrome, primary biliary cirrhosis, polymyositis, and autoimmune hepatitis The method according to any one of claims 13 to 19, which is a selected disease.
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US9856278B2 (en) | 2014-07-04 | 2018-01-02 | The University Of Tokyo | Lysophosphatidylserine derivative |
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WO2023145873A1 (en) * | 2022-01-27 | 2023-08-03 | 国立大学法人東京大学 | Lysophosphatidylserine analogue, and lysophosphatidylserine analogue-containing phamaceutical composition for treating or preventing fibrosis |
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