WO2012136121A1

WO2012136121A1 - Amino phenylpropanoid compound and use thereof for preparing immunosuppressant

Info

Publication number: WO2012136121A1
Application number: PCT/CN2012/073336
Authority: WO
Inventors: 李松; 王晓奎; 陈耀; 钟武; 刘洪英; 郑志兵; 谢云德; 赵国明
Original assignee: 中国人民解放军军事医学科学院毒物药物研究所
Priority date: 2011-04-02
Filing date: 2012-03-30
Publication date: 2012-10-11
Also published as: CN103492359B; CN103492359A

Abstract

An amino phenylpropanoid compound, or a geometric isomer thereof, or a pharmaceutically acceptable salt or hydrate thereof as represented by Formula I, a preparation method therefor, and a pharmaceutical composition containing the compound. The compound is used to act as an immunosuppressant, applicable in preparing an organ transplant anti-rejection medicament and medicaments for treating and/or preventing certain autoimmune diseases such as rheumatoid, psoriasis, multiple sclerosis, and systemic lupus erythematosus. [Formula I]

Description

Aminophenylpropanol compounds and their use

Use of preparation of immunosuppressants

The present invention relates to an aminophenylpropanol compound or a geometric isomer thereof or a pharmaceutically acceptable salt or hydrate thereof, a process for producing the same, and a pharmaceutical composition containing the compound. The present invention also relates to the use of the compound as an immunosuppressive agent for the preparation of a medicament for combating organ transplant rejection and for treating and/or preventing certain autoimmune diseases such as rheumatoid, psoriasis, multiple sclerosis, systemic lupus erythematosus and the like. the use of. Background technique

The present invention relates to a class of compounds having immunomodulatory effects and their use as pharmaceutical and/or pharmaceutical compositions.

Immune disorders can lead to a range of autoimmune diseases and chronic inflammatory diseases such as systemic lupus erythematosus, chronic rheumatoid arthritis, type I and type II diabetes, certain enteritis, biliary cirrhosis, multiple sclerosis and other diseases, such as Crohn's colitis, ulcerative colitis, large pemphigus, sarcomatoid disease and asthma, and so on. Although the pathogenesis and processes of these diseases are different, they are identical in terms of autoantibody production and lymphocyte self-reaction. It may be because the normal immune system balance is broken. Similarly, after bone marrow or organ transplantation, the host's lymphocytes recognize foreign tissue antigens and begin to produce cellular and humoral responses, including antibodies, cytokines, and cytotoxic lymphocytes, leading to transplant rejection. It is also likely that the normal immune system balance is broken.

One of the end result of autoimmune diseases and transplant rejection is that inflammatory cells and their released regulators cause tissue damage. Although many immunosuppressants have been used clinically, they each have different disadvantages. For example, early, immunosuppression The preparation is represented by glucocorticoids and azathioprine. The combined use of the two made the first breakthrough in organ transplantation, but until the end of the 1970s, the combined use of the two as a kidney transplant to maintain immunosuppressive therapy had a one-year survival rate of only about 50%. At the same time, long-term application of glucocorticoids can cause serious adverse reactions, such as inducing and aggravating infections, leading to adrenal cortical dysfunction, etc. Long-term use of cytotoxic immunoassays such as azathioprine leads to inhibition of bone marrow hematopoietic system and gastrointestinal mucosa damage. Subsequently, the application of cyclosporine A (CsA) has made great progress in the transplantation of various organ transplants, including liver, heart, cardiopulmonary, pancreas, and bone marrow. Until the mid-1980s, most immunosuppressive treatment regimens were still based on cyclosporine A, but cyclosporin A had significant toxicity, such as nephrotoxicity, hepatotoxicity, hyperglycemia, neurological damage, infection, and tumorigenesis. . Mycophenolate mofetil is an ester derivative of mycophenolic acid and has a strong immunosuppressive effect. In 1996, clinical trials showed that the mycophenolate mofetil group reduced the incidence of acute renal allograft rejection by nearly 50% compared with the combination of mycophenolate mofetil, prednisone and cyclosporine A, and significantly reduced the rate of renal transplantation. Failure rate. At the same time, mycophenolate mofetil can reverse the vascular rejection that is not effective in conventional immunosuppressive therapy, thereby avoiding graft failure. Mycophenolate mofetil has lower hepatotoxicity, nephrotoxicity and myelosuppression, and no side effects such as hypertension, diabetes, pancreatitis, osteoporosis, etc. However, some of its toxic reactions in clinical applications are still unbearable. The discovery of FK506 has brought organ transplantation into another new era. Compared with the cyclosporine A treatment group, the survival rate of the FK506 treatment group was significantly increased, and the rejection occurred less, and the amount of cholesterol-like drugs was also reduced. The literature also showed that after the application of cyclosporine A, Rejection, 65%-80% can be reversed by FK506. Furthermore, FK506 has the advantage that its proliferative effect on hepatocytes makes it more beneficial for liver transplantation and does not produce side effects of some tissue proliferation caused by cyclosporine A. Relatively speaking, the application of FK506 has a high survival rate of transplant patients and a high graft survival rate, a low incidence of acute rejection, a relatively low dependence on cholesterol-like, and a low infection rate. However, FK506 still has side effects such as nephrotoxicity, neurotoxicity, tumors, infections, allergic reactions, high blood pressure and high blood sugar. It can be seen that the existing immunosuppressive agents can not meet the needs of clinical treatment, and it is still necessary to find new high-efficiency and low-toxic immunosuppressive agents.

Accordingly, it is an object of the present invention to provide a safer, more effective immunosuppressive active compound. Summary of the invention

The inventors have found through research that the compound represented by the formula I has an immunosuppressive action. It can be used for, but not limited to, anti-immunological rejection in organ transplantation, prevention and/or treatment of certain autoimmune diseases such as rheumatoid, psoriasis, multiple sclerosis, systemic lupus erythematosus, Crohn's disease, ulcerative large intestine Inflammation, major pemphigus, rickets, and asthma.

Accordingly, one aspect of the invention relates to an aminophenylpropanol compound of the formula I or a geometric isomer thereof or pharmaceutically acceptable

among them-.

n is 1, 2 or 3, preferably 1,

X is 0, S, NH or Se, etc., preferably 0,

Is hydrogen, an alkyl group or an acyl group having a carbon number of 1-8, preferably 1-4,

R ₂ is an alkyl or acyl substituent having a carbon number of 2 to 20, preferably 6 to 12, and the alkyl or acyl substituent may or may not contain an aromatic substituent, and each aromatic substituent may be 1 to 5 halogens. Substituted, wherein the alkyl or acyl substituent may contain from 1 to 3 sulfur, nitrogen or an oxygen atom to the alkyl group and the acyl group, the alkyl group and the aromatic substituent, the acyl group Aromatic substituents, or different or identical aromatic substituents, and

R ₃ is a hydrogen atom, an alkyl group having an C number of 1-8, preferably 1-4, an acyl group or an alkoxy group. The invention is further described as a compound of formula la or a geometric isomer or pharmaceutically acceptable salt or hydrate thereof:

R ₂ is an alkyl or acyl substituent having a carbon number of 2 to 20, preferably 6 to 12, and the alkyl or acyl substituent may or may not contain an aromatic substituent, and each aromatic substituent may be 1 to 5 halogens. Substituted, wherein the alkyl or acyl substituent may contain from 1 to 3 sulfur, nitrogen or an oxygen atom to the alkyl group and the acyl group, the alkyl group and the aromatic substituent, the acyl group and the aromatic substituent, or a different or identical aromatic substituent, with

R ₃ is a hydrogen atom, an alkanoyl group having a carbon number of 1-8, preferably 1-4 or an alkoxy group. The invention is further described as a compound of formula lb or a geometric isomer or pharmaceutically acceptable salt or hydrate thereof:

R ₂ is an alkyl or acyl substituent having a carbon number of 2 to 20, preferably 6 to 12, the alkyl group Or an acyl substituent, which may or may not contain an aromatic substituent, each aromatic substituent may be substituted with from 1 to 5 halogens, wherein the alkyl or acyl substituent may contain from 1 to 3 sulfur, nitrogen or an oxygen atom to the alkyl group. And an acyl group, an alkyl group and an aromatic substituent, an acyl group and an aromatic substituent, or a different or the same aromatic substituent;

R ₃ is a hydrogen atom, an alkanoyl group having a carbon number of 1-8, preferably 1-4 or an alkoxy group. The term "aromatic substituent" as used herein means a carbocyclic or heterocyclic aromatic ring group. The carbocyclic aromatic ring group is a group having a single ring or a plurality of fused rings having a carbon number of 5 to 22, preferably 6 to 18, 6 to 14, 6 to 10, more preferably 6. Typical examples of the carbocyclic aromatic ring group include, but are not limited to, a phenyl group, a naphthyl group, an anthracenyl group and the like. The heterocyclic aromatic ring group is a group having 5-14, preferably 5-10 members having one single ring or a plurality of fused rings having one or more hetero atoms selected from N, 0 and S. Typical examples of the heterocyclic aromatic ring group include, but are not limited to, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazole , isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, benzimidazolyl, benzothienyl, benzofuranyl, fluorenyl, benzotrien Azyl, benzothienyl, benzoxazolyl, isoquinolinyl, fluorenyl, isodecyl, acridinyl, benzisoxazolyl, fluorenyl, pyrazinyl, quinolyl And isoquinolinyl and the like.

The term "halogen" as used herein means fluoro, chloro, bromo or iodo. Preferred halogens are fluorine, chlorine or bromine, more preferably fluorine or bromine. The compounds of formula I according to the invention are preferably, but not limited to, the following compounds:

Another aspect of the invention relates to a pharmaceutical composition comprising at least one A compound of formula I or a geometric isomer or pharmaceutically acceptable salt or hydrate thereof and one or more pharmaceutically acceptable carriers or excipients.

A further aspect of the invention relates to a process for the preparation of a compound of formula lb, or a pharmaceutically acceptable salt or hydrate thereof, specifically comprising:

1. Synthesis of (S)-2-amino-3-(4-acylphenyl)-propan-1-ol

R=C _n H _2n+1 , n=5-ll This method uses L-phenylalanine as a raw material, and reduces carboxylic acid with NaBH ₄ /I ₂ to obtain L-phenylalaninol, reacetylation, Fu- A five-step reaction such as acylation, deprotection, and salt formation gives the target compound.

2. Synthesis of (S)-2-amino-3-(4-alkylphenyl)-propan-1-ol compound This route uses L-phenylalanine as a raw material to reduce carboxylic acid with NaBH ₄ /I ₂ The target compound is obtained by a six-step reaction such as L-phenylalaninol, reacetylation, Friedel-Craft acylation, reduction of Et ₃ SiH/TFA system, deprotection, and salt formation. The reduction, acetylation and Friedel-Craft acylation methods are the same as in 2.

3. Synthesis of (S)-2-amino-3-(4-substituted benzyloxy-phenyl)-propan-1-ol

The route uses L-Boc tyrosine methyl ester as a raw material, and the target compound is obtained by a benzyl group protecting phenolic hydroxyl group, LiAlH ₄ reduction, deprotection, alkylation, de Boc reaction and the like.

4. Synthesis of (S)-2-amino-3-(4-substituted phenoxy-acetylphenyl)-propan-1-ol compound

This route uses L-phenylalanine as a raw material, and reduces carboxylic acid with NaBH ₄ /I ₂ to obtain L- Phenylalanin, reacetylation, chloroacetylation, condensation with phenol, deprotection, salt formation, etc., to obtain the target compound.

R = Br A pharmaceutically acceptable salt of a compound of the invention includes an acid addition salt thereof with an inorganic or organic acid or a base addition salt with a base. The acid addition salts include, but are not limited to: hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, acetate, propionate, butyl Acid salt, trimethylacetate, adipate, alginate, lactate, citrate, tartrate, succinate, maleate, fumarate, picrate, winter Alkanoate, gluconate, benzoate, methanesulfonate, ethanesulfonate, besylate, p-toluenesulfonate and pamoate; base addition salts include but are not limited to: Ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, organic base salts such as dicyclohexylamine and N-methyl-D-glucosamine salts, and ammonia

.

The pharmaceutical composition of the compound of the present invention can be administered in any of the following ways: oral, spray inhalation, rectal administration, nasal administration, buccal administration, vaginal administration, topical administration, parenteral administration such as subcutaneous, intravenous, intramuscular, intraperitoneal Intrathecal, intraventricular, intrasternal, and intracranial injection or input, or by means of an explant reservoir. Among them, oral administration, intraperitoneal or intravenous administration and topical administration are preferred. When administered orally, the compounds of the invention may be formulated in any orally acceptable form including, but not limited to, tablets, capsules, aqueous solutions or aqueous suspensions. Among them, carriers which are generally used for tablets include lactose and corn starch, and a lubricant such as magnesium stearate may also be added. Diluents commonly used in capsule preparations include lactose and dried corn starch. Aqueous suspension formulations are usually prepared by admixing the active ingredient with a suitable emulsifier and suspension. If desired, some sweeteners, fragrances or colorants may be added to the above oral formulations.

When administered rectally, the compounds of the invention will generally be in the form of a suppository prepared by admixing the drug with a suitable non-irritating excipient. The excipient exhibited a solid state at room temperature and melted at a rectal temperature to release the drug. Such excipients include cocoa butter, beeswax and polyethylene glycol.

When applied topically, especially in the treatment of facial or organs easily accessible by topical application, such as eye, skin or lower intestinal neurological diseases, the compounds of the present invention can be formulated into different topical preparations according to different affected faces or organs. Specifically, the following description is as follows: When the eye is applied topically, the compound of the present invention can be formulated into a preparation form of a micronized suspension or solution, and the carrier used is an isotonic pH of sterile saline, which may or may not be preserved. An agent such as a benzyl alkoxide is chlorinated. In addition, for ophthalmic use, the compound can also be formulated into a cream form such as a vaseline cream.

When applied topically to the skin, the compounds of the invention may be in the form of a suitable ointment, lotion or cream preparation wherein the active ingredient is suspended or dissolved in one or more carriers. Carriers which may be used herein for ointment preparations include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide, emulsifying wax and water; and lotions which may be used for lotions or creams include, but are not limited to: Mineral oil, sorbitan monostearate, Tween 60, cetyl esters wax, hexadecene aryl alcohol, 2-octyldodecanol, benzyl alcohol and water.

When applied topically to the lower intestinal tract, the compounds of the invention may be formulated into the rectum as described above In the form of a suppository formulation or a suitable enteral preparation, a topical transdermal patch may also be employed. The compounds of the present invention can also be administered in the form of a sterile injectable preparation, including sterile injectable aqueous or oily suspensions, or sterile injectable solutions. Among them, carriers and solvents which can be used include water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils may be employed as a solvent or suspension medium such as a monoglyceride or a diglyceride.

In addition, the compounds of the invention may be administered with other immunosuppressive drugs, including but not limited to: cyclosporin A, steroid hormones, FK506, RPM, Lef lunomide, DSG, SKF105685MZ, RS61443BQR, etc.

It should also be noted that the specific dosage and method of use of the compounds of the present invention for different patients depends on a number of factors, including the age, weight, sex, natural health, nutritional status of the compound, the strength of the compound, the time of administration, the rate of metabolism, the condition. The severity of the diagnosis and the subjective judgment of the doctor. A preferred dose is between 0.1 and 300 mg/kg body weight per day. Example

The following examples are intended to describe the invention in further detail, but without restricting the invention.

The melting point of the compound was determined by a RY-1 type melting point apparatus, and the thermometer was not corrected. - NMR was measured by an ARX-400 NMR instrument. The quality was measured by a Micromas s-ZabSpec MS instrument. All reactions were pretreated with a solvent that was not indicated.

Example 1 : Preparation of (S)-2-amino-3-phenyl-propan-1-ol

In a 1 L three-necked flask, 6.92 g (183 leg ol) of sodium borohydride and 200 mL of anhydrous tetrahydrofuran were placed, and the mixture was cooled in a water bath, and 12.56 g (76 mmol) of L-phenylalanine was added in portions. 5小时后旋溶剂。 Then, after the addition of 19.30g (76 lb) of iodine in anhydrous tetrahydrofuran solution 80mL, 30min, and the mixture was heated to reflux for 18h, then cooled to room temperature, adding methanol to the reaction liquid was clarified, stirring at room temperature 0. 5h, solvent was removed, then Add 20% potassium hydroxide The solution was stirred with aq. EtOAc (EtOAc EtOAc. 91-93 ) ₀ ¹ H-NMR (400 MHz, CDC1 ₃ ) δ (ppm): 2.92 (d, 2H), 3.40-3.51 (m, 2H), 3.65-3.7 (q, 1H), 7.25-7.32 (m, 5H).

Example 2: Preparation of (S)-2-ethylamino-3-phenyl-propan-1-ol acetate In a 250 mL round bottom flask, 19.66 g (130 mmol) of the compound of Example 1, 132.72 g ( l. 3mol) acetic anhydride, cooled to O in a water bath, dripping 132 drops of sulfuric acid, naturally warmed to room temperature overnight, slowly poured into 300mL of saturated sodium carbonate solution, a large amount of white solid precipitated, filtered, washed with a suitable amount of water, and dried in vacuo to give 26.3g of white solid, a yield of 86.0% "^ -NMR (400 MHz , CDC1 3) δ (ppm): 1.96 (s, 3H), 2.10 (s, 3H), 2.82-2.89 (m, 2H ), 4.04-4.07 (m, 2H), 4.44 (m, 1H), 5.62 (d, 1H), 7.20-7.31 (m, 5H).

Example 3: Preparation of (S)- 2-acetamido-3-(4-octanoylphenyl)-propan-1-ol acetate

The compound obtained in Example 2 (4.71 g, 20 mmol) was added to 50 mL of 1,2-dichloroethane, cooled to 0, anhydrous aluminum trichloride (5.32 g, 40 mmol) was added, and octanoyl chloride (5.85 g) was added dropwise. , 36mmol) 1,2-dichloroethane 20mL, add, heat the reaction for 4 hours, then stir at room temperature for 2 hours, the reaction solution was poured into crushed water, shaken, liquid, water phase with dichloromethane 50x3 The organic phase was combined, washed with EtOAc EtOAc EtOAc EtOAc. ¹ H-NMR (400 MHz, CDC1 ₃ ) δ (ppm): 0.88 (t, 3H), 1.29-1.34 (m, 8Η), 1.70-1.74 (m, 2Η), 1.97 (s, 3Η), 2.11 ( s, 3Η), 2.90-2.96 (m, 4Η), 4.07 (m, 2Η), 4.46 (m, 1Η), 5.67 (d, 1H), 7.28-7.30 (d, J = 8.0Hz, 2H), 7.89 -7.91 (d, J = 8.0Hz, 2H). Example 4: Preparation of (S)-2-amino-3-(4-octanoylphenyl)-propan-1-ol In a 100 mL round bottom flask, 0.36 g (1.0 mmol) of the compound of Example 3 was added. 0.42g (10 legs. 1) lithium hydroxide, 30mL of methanol, 30mL of distilled water, heated under reflux for 7h, extracted with ethyl acetate 50mLx 3 extraction, combined, washed once with saturated sodium chloride solution, dried anhydrous sodium sulfate, filtered, It was dried to dryness, and ethyl acetate was recrystallized to give 0.25 g of white solid.

Example 5: Preparation of (S)- 2-amino-3-(4-octanoylphenyl)-propan-1-ol hydrochloride

The compound of Example 4 (0.14 g, 0.5 mmol) was added to 5 mL of ethyl acetate to dissolve it, and hydrogen chloride (2.2 mol. L ¹ ) in anhydrous ethyl acetate solution was added 1. OmL, stirred at room temperature for 2 hours, with white The solid was precipitated, the ethyl acetate was evaporated under reduced pressure, and then crystallised from ethyl acetate/ethanol to give crystals of white crystals. llg, yield 73.0%, mp 107-108X _o ¹ H-NMR (400 MHz, D ₂ 0) δ (ppm): 0.68-0.72 (t, 3H), 1.11-1.17 (m, 8H), 1.52 (m, 2H), 2.92 (m, 4H), 3.52 (m, 2H), 3.67 (m, 1H), 7.31-7.33 (d, J = 8. OHz, 2H), 7.83-7.85 (d, J = 8.0 Hz, 2H).

Example 6: Preparation of (S)- 2-amino-3-(4-hexanoylphenyl)-propan-1-ol hydrochloride

The title compound was obtained by the method of Example 5 (yield: 7). ^- MR (400 MHz, CD ₃ 0D) δ (ppm): 0.92 (t, 3H), 1.37 (m, 4Η), 1.67-1.69 (m, 2Η), 3.01 (m, 4Η), 3.50 (m, 2Η), 3.70 (m, 1Η), 7.42-7.44 (d, J = 8.0Hz, 2H), 7.97-7.99 (d, J = 8. OHz, 2H).

Example 7: Preparation of (S)- 2-amino-3-(4-heptanoylphenyl)-propan-1-ol hydrochloride

Using (S)-2-amino-3-(4-heptanoylphenyl)-propan-1-ol as raw material, using examples 5 Method The title compound was obtained in a yield of 71.30%. - Legs (400 MHz, CD ₃ 0D) δ (ppm): 0.90 (t, 3H), 1.32-1.33 (m, 6H), 1.64-1.70 (m, 2H), 2.98-3.02 (m, 4H), 3.48 -3.51 (m, 2H), 3.68 (m, 1H), 7.40-7.42 (d, J = 8.0 Hz, 2H), 7.96-7.98 (d, J = 8.0 Hz, 2H).

Example 8: Preparation of (S)- 2-amino-3-(4-nonanoylphenyl)-propan-1-ol hydrochloride

Starting from (S)-2-amino-3-(4-nonanoylphenyl)-propan-1-ol, the title compound was obtained by the procedure of Example 5 to yield 69.65%. - Legs (400 MHz, CD ₃ 0D) δ (ppm): 0.87- 0.90 (t, 3H), 1.28-1.31 (m, 10H), 1.66- 1.70 (m, 2H), 2.98-3.02 (m, 4H) , 3.48-3.51 (m, 2H), 3.68 (m, 1H), 7.40-7.42 (d, J = 8.0 Hz, 2H), 7.96-7.98 (d, J = 8·0Ηζ, 2H).

Example 9: Preparation of (S)- 2-amino-3-(4-nonanoylphenyl)-propan-1-ol hydrochloride

Starting from (S)-2-amino-3-(4-nonanoylphenyl)-propan-1-ol, the title compound was obtained by the procedure of Example 5 in a yield of 65.45%. ^- MR (400 MHz, CD ₃ 0D) δ (ppm): 0.87-0.91 (t, 3H), 1.28-1.35 (m, 12H), 1.67- 1.69 (m, 2Η), 2.99-3.00 (m, 4Η) ), 3.48-3.51 (m, 2Η), 3.70 (m, 1Η), 7.41-7.43 (d, J = 8.0Hz, 2H), 7.97-7.99 (d, J = 8·0Ηζ, 2H).

Example 10: Preparation of (S)- 2-amino-3-(4-n-undecylphenyl)-propan-1-ol hydrochloride

The title compound was obtained by the method of Example 5 using (S)-2-amino-3-(4-n-undecylphenyl)-propan-1-ol as the starting material, yield 70.19% „ ^-NMR (400 MHz , CD ₃ 0D) δ (ppm): 0.86-0.90 (t, 3H), 1.26- 1.38 (m, 14H), 1.66- 1.70 (m, 2Η), 2.98-3.01 (m, 4Η), 3.49-3.50 ( m, 2Η), 3.71(m, 1Η), 7.40-7.42 (d, J = 8.0Hz, 2H), 7.96-7.98 (d, J = 8·0Ηζ, 2H).

Example 11: (S)- 2-Amino-3-(4-n-dodecanoylphenyl)-propan-1-olate Preparation of acid salt

To (S) -2- amino - 3- (4-n-dodecanoyl-phenyl) - propan --1- alcohol as raw materials, to give the title compound Example 5, a yield of 59.84% ₀ ^ -NMR (400 MHz , CD ₃ 0D) δ (ppm): 0.86-0.90 (t, 3H), 1.27- 1.36 (m, 16H), 1.66- 1.70 (m, 2H), 2.98-3.01 (m, 4H), 3.49-3.52 ( m, 2H), 3.71(m, 1H), 7.40-7.42 (d, J = 8.0Hz, 2H), 7.96-7.98 (d, J = 8·0Ηζ, 2H).

Example 12: Preparation of (S)- 2-amino-3-(4-octylphenyl)-propan-1-ol hydrochloride

In a 50 mL round bottom flask, 0.83 g (2.3 mmol) of the compound of Example 3, 5. OmL TFA was added, cooled in a water bath, and 0.80 mL of triethylsilane was slowly added dropwise, and the mixture was stirred at room temperature overnight, and poured into 10 mL of saturated The aqueous solution of sodium carbonate was extracted with ethyl acetate (50 mL), and the mixture was combined, washed with EtOAc EtOAc EtOAc EtOAc EtOAc %.

In a 100 mL round bottom flask, 0.40 g (1.2 mmol) of the above compound, 0.50 g (12 leg ol) of lithium hydroxide, 30 mL of methanol, 30 mL of distilled water were added, and the mixture was heated under reflux for 7 h, extracted with ethyl acetate (50 mL×3), and combined. washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, spin-dry, a white solid was recrystallized from ethyl acetate to give 0.31 g, yield 87.0% ₀

The above compound (0.31 g, 1.2 leg ol) was added to 5 mL of ethyl acetate to dissolve it, and hydrogen chloride (2.2 mol. L - anhydrous ethyl acetate solution 1.0 mL) was added thereto, and the mixture was stirred at room temperature for 2 hours, and a white solid was precipitated. , ethyl acetate was removed under reduced pressure, to give 0.25g of white flaky crystals from ethyl acetate / recrystallized from ethanol, yield 71.0%, mp 107-108X NMR (400 MHz, CD 3 0D) δ (ppm):. 0.87- 0.90 (t, 3H), 1.28-1.31 (m, 10H), 1.57-1.60 (m, 2H), 2.57-2.59 (t, 2H), 2.88-2.90 (m, 2H), 3.42-3.45 (m, 1H) ), 3.50-3.53 (q, 1H), 3.67-3.69 (q, IH), 7.17(s, 4H).

Example 13: Preparation of (S)- 2-amino-3-(4-hexylphenyl)-propan-1-ol hydrochloride

Starting from (S)-2-acetamido-3-(4-hexanoylphenyl)-propan-1-ol acetate, the title compound was obtained from m. - NMR (400 MHz, CD ₃ 0D) δ (ppm): 0.87-0.90 (t, 3H), 1.30-1.31 (m, 6H), 1.57-1.60 (m, 2H), 2.56-2.60 (t, 2H) , 2.90-2.91 (m, 2Η), 3.45 (m, IH), 3.50-3.54 (q, IH), 3.66-3.69 (q, IH), 7.17(s, 4H).

Example 14: Preparation of (S)- 2-amino-3-(4-heptylphenyl)-propan-1-ol hydrochloride

Starting from (S)-2-acetamido-3-(4-heptanoylphenyl)-propan-1-ol acetate, the title compound was obtained from m. - NMR (400 MHz, CD3OD) δ (ppm): 0.87-0.90 (t, 3H), 1.30-1.32 (m, 8H), 1.59-1.61 (m, 2H), 2.57-2.59 (t, 2H), 2.88 -2.90 (m, 2Η), 3.41-3.45 (m, IH), 3.50-3.54 (q, IH), 3.67-3.69 (q, IH), 7.17 (s, 4H).

Example 15: Preparation of (S)- 2-amino-3-(4-mercaptophenyl)-propan-1-ol hydrochloride

Starting from (S)-2-acetamido-3-(4-indoleylphenyl)-propan-1-ol acetate, the title compound was obtained by the method of Example 12, yield 66.42%. - NMR (400 MHz, CD3OD) δ (ppm): 0.87-0.90 (t, 3H), 1.28-1.31 (m, 12H), 1.59 (m, 2H), 2.57-2.61 (t, 2H), 2.87-2.89 (d, 2H), 3.42-3.46 (m, IH), 3.49-3.52 (q, IH), 3.65-3.70 (q, IH), 7.17(s, 4H).

Example 16: Preparation of (S)- 2-amino-3-(4-mercaptophenyl)-propan-1-ol hydrochloride

Taking (S)-2-acetamido-3-(4-nonanoylphenyl)-propan-1-ol acetate as the original The title compound was obtained by the method of Example 12 to yield 65.88%. ^- MR (400 MHz, CD ₃ 0D) δ (ppm): 0.87-0.91 (t, 3H), 1.27-1.31 (m, 14H), 1.57-1.59 (m, 2H), 2.57-2.58 (t , 2H ), 2.88-2.90 (d, 2Η), 3.42-3.45 (m, 1Η), 3.49-3.51 (q, 1H), 3.66-3.69 (q, 1H), 7.17 (s, 4H).

Example 17: Preparation of (S)- 2-amino-3-(4-n-undecylphenyl)-propan-1-ol hydrochloride

Starting from (S)-2-acetamido-3-(4-n-undecylphenyl)-propan-1-ol acetate, the title compound was obtained by the method of Example 12, yield 56.66%. 'H-NMR (400 MHz, CD ₃ OD) δ (ppm): 0.87-0.91 (t, 3H), 1.27-1.31 (m, 16H), 1.57-1.61 (m, 2H), 2.57-2.61 (t, 2H), 2.88-2.90 (d, 2H), 3.41-3.44 (m, 1H), 3.48-3.50 (q, 1H), 3.66-3.69 (q, 1H), 7.17(s, 4H).

Example 18: Preparation of (S)- 2-amino-3-(4-n-dodecylphenyl)-propan-1-ol hydrochloride

Starting from (S)-2-acetamido-3-(4-n-dodecanoylphenyl)-propan-1-ol acetate, the title compound was obtained from m. 'H-NMR (400 MHz, CD ₃ OD) δ (ppm): 0.87-0.91 (t, 3H), 1.27-1.31 (m, 18H), 1.57-1.61 (m, 2H), 2.57-2.60 (t, 2H), 2.88-2.90 (d, 2H), 3.42-3.45 (m, 1H), 3.48-3.50 (q, 1H), 3.66-3.70 (q, 1H), 7.17(s, 4H).

Example 19: Preparation of (S)-3-(4-benzyloxy-phenyl)-2-tert-butoxycarbonylamino-propan-1-ol

In a 500 mL round bottom flask, 16.0 g (54.2 leg ol) of (S)-3-(4-hydroxyphenyl)-2-tert-butoxycarbonylamino-propan-1-carboxylate, 10.19 g ( 59.6 leg ol) benzyl bromide, 22.4g (162.6mmol) carbonic acid clock, 200mL acetone, heated to reflux to the raw material After disappearing, filtration, the cake was washed with 50 mL of ethyl acetate, and evaporated to ethyl ether.

To a 250 mL three-necked flask, 0.34 g (8.9 mmol) of lithium aluminum tetrahydrochloride and 10 mL of anhydrous tetrahydrofuran were added, and heated to dropwise add 4.26 g (11. mmol) of 40 mL of the above-mentioned compound anhydrous tetrahydrofuran solution, and the reaction was kept overnight. Slowly pour into 100 mL of water, separate the organic layer, add 10% sulfuric acid to the aqueous layer until the precipitate disappears, extract 100 mL×3 with ethyl acetate, combine, wash once with saturated sodium chloride solution, dry with anhydrous sodium sulfate. The lg white solid, yield 78.0%, EtOAc (EtOAc:EtOAc: ¹ H-NMR (400 MHz, DMSO-^) δ (ppm): 1.28-132 (s, 9H), 2.47-2.51 (q, 1Η), 2.71-2.75 (q, 1Η), 3.24-3.34 (m, 2H), 3.50 (m, 1H), 4.66-4.69 (t, 1H), 5.05 (s, 2H), 6.56-6.58 (d, 1H), 6.89-6.91 (d, J = 8.0Hz, 2H), 6.89 -6.91 (d, J = 8.0 Hz, 2H), 7.09-7.11 (d, J = 8.0 Hz, 2H), 7.32-7.429 (m, 5H). ESI-MS m/z (M+H+) 358.13.

The compound (S)-3-(4-benzyloxy-phenyl)-2-tert-butoxycarbonylamino-propan-1-ol is refluxed in trifluoroacetic acid to give (S)-3-(4) -benzyloxy-phenyl)-2-amino-propan-1-ol trifluoroacetate.

ESI-MS m/z (M+H ⁺ ) 371.16.

Example 20: Preparation of (S)-3-[4-(3,4-difluoro-benzyloxy)-phenyl]-2-amino-propan-1-ol

In a 250 mL round bottom flask, 7.15 g (20 lb) of the compound of Example 19, 7.63 g (120 lb) of ammonium formate, 0.72 g of palladium carbon, 200 mL of absolute ethanol were added, and the mixture was heated to reflux 4. Oh, and the palladium carbon was recovered by filtration. , spin dry to give 4.38 g of solid, yield 82.0%.

In a lOOmL round bottom flask, 0.89 g (3.0 lb ol) of the above compound, 0.58 g (3.6 mmol) of 3,4-difluoro-benzyl chloride, 1.66 g (12-leg ol) potassium carbonate, 0.1 g (0.6 g) Methyl) potassium iodide, 50mL of acetone, heated to reflux until the starting material disappears, filtered, The filter cake was washed with 10 mL of ethyl acetate and dried and evaporated. Yield 60.0% ₀ ^-NMR (400 MHz, CD ₃ 0D) δ (ppm): 2.56-2.61 (q, 1H), 2.77-2.82 (q, 1H), 3.44-3.52 (m, 2H), 4.00- 4.03 (m, 1H), 4.92 (s, 2H), 6.80-6.82 (d, J = 8.0Hz, 2H), 7.03-7.05 (d, J = 8.0Hz, 2H), 7.12— 7.27 (m, 3H) .

Example 21: Preparation of (S)-3-[4-(2,6-difluoro-benzyloxy)-phenyl]-2-amino-propan-1-ol

The title compound was obtained by the method of Example 20 using 2, 6-difluorobenzyl chloride instead of 3, 4-difluorobenzyl chloride as a starting material, yield 58.0%. - Leg (400 MHz, CD ₃ 0D) δ (ppm): 2.67-2.72 (q, 1H), 2.87-2.92 (q, 1H), 3.54-3.62 (m, 2H), 4.10-4.12 (m, 1H) , 5.08 (s, 2H), 6.88-6.90 (d, J = 8.0Hz, 2H), 6.98-7.05 (m , 2H) , 7.13-7.15 (d , J = 8.0Hz , 2H) , 7.38-7.44 (m , 1H).

Example 22: Preparation of (S)-2-ethylamino-3-(4-chloroacetylphenyl)-propan-1-ol acetate

The compound of Example 2 (9.4 g, 40 leg ol) was added to 100 mL of 1,2-dichloroethane, and the temperature was lowered until anhydrous aluminum trichloride (17.07 g, 128 mmol) was added, and chloroacetyl chloride (5.42 g) was added dropwise. , 48 leg ol) 1,2-dichloroethane 20mL, add, heat the reaction for 4 hours, then stir at room temperature for 2 hours, the reaction solution was poured into crushed water, shaken, liquid, the aqueous phase with acetic acid The ester was extracted with 100 mL of 3, and the organic phase was combined, washed twice with water, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and dried to give 9.35 g of pale yellow solid, yield 73.0% „ ^-NMR (400 MHz, CDC1 ₃ ) δ (ppm): 1.97 (s, 3H), 2.11 (s, 3H), 2.87-2.98 (m, 2H), 4.03-4.13 (m, 2H), 4.47 (m, 1H), 4.71 (s, 2H), 5.70-5.73 (d, 1H), 7.34-7.36 (d, J = 8.0Hz, 2H), 7.90-7.92 (d, J = 8.0Hz, 2H).

Example 23: (S)-2-Amino-3- [4-(4-bromo-phenoxy-acetyl)-benzene Preparation of -propan-1-ol

In a lOOmL round bottom flask, 1.56 g (5.0 mmol) of the compound of Example 22, 1.72 g (6 mmol) of p-bromophenol, 2.07 g (15 mmol) of potassium carbonate, 0.1 g (0.5 mmol) of potassium iodide, 80 mL of acetone, and heated reflux were added. After 4 h, the mixture was filtered, washed with EtOAc EtOAc EtOAc EtOAc EtOAc

In a 250 mL round bottom flask, 1.77 g (4.0 lb ol) of the above compound, 1.68 g (40 mmol) of lithium hydroxide, 80 mL of methanol, and 80 mL of distilled water were added, and the mixture was heated to reflux for 7 h, cooled to room temperature, and methanol was evaporated. The extract was extracted with 100 mL of EtOAc (3 mL), EtOAc (EtOAc) ^- MR (400 MHz, DMS0-) δ (ppm): 1.36 (s, 2H), 2.48-2.51 (q, 1H), 2.76-2.81 (q, 1H), 2.89-2.92 (m, 1H), 3.21 -3.36 (m, 2H), 4.66 (s, 1H), 5.58 (s, 2H), 6.93-6.96 (m, 2H), 7.40-7.46 (m, 4H), 7.92-7.94 (m, 2H).

Example 24: Preparation of (S)-3-[4-(4-decyloxy-benzyloxy)-phenyl]-2-amino-propan-1-ol trifluoroacetate

The experimental procedure was the same as in Example 19, using the compound (S)-3-(4-hydroxyphenyl)-2-tert-butoxycarbonylamino-propano-1-methyl ester and 1-(bromomethyl)-4. -Methoxybenzene is used as a raw material to obtain a product. Yield: 75.5%.

ESI-MS m/z (M+H ⁺ ) 401.14.

Example 25: Screening of Inventive Examples 5 - 21 and 23 - 24 Compounds Using the EGFP-S1P1-U20S Cell Model

1. Materials and instruments:

1.1 Instruments: IN Cell Analyzer 1000 Live Cell Imaging System (GE, USA) 1. 2 cell line: U20S expressing EGFP-S1P1 fusion protein (a subsidiary of Thermom Biolmage, USA)

1. 3 reagent preparation:

Cell culture medium: DMEM high glucose medium containing 0.5 mg/ml G418 and 10% FBS

Analytical culture solution: F12 medium containing l OmM HEPES, 0.1% defatting BSA 3 Fixative: 12% formaldehyde solution (PBS)

Staining solution: PBS containing Ι μ Μ Hoechs t 33342 (Invitrogen, USA)

1. 4 Preparation of the compound:

Including 4%. The assay medium of DMS0 was used as a 4X control working solution with a final concentration of 1% DMS0.

The agonist sphingosine-1-phosphate S1P (natural ligand, Sigma, MW=379. 5, EC ₅ . ~ 25 nM, sometimes referred to as S1P hereinafter) is formulated into a 3 mM mother liquor with 1 OmM NaOH aqueous solution. 4%. The analytical medium of DMS0 was formulated into 4 χ working solution and given final concentrations of 10 nM, 250 nM and 1 μ Μ.

Inventive Examples 5 - 21 and 23 - 24 Compounds were formulated into 30 mM mother liquors using DMS0, diluted to a continuous concentration gradient of 1000 X mother liquor using DMS 0 times, and then analyzed to prepare 4 working solutions. Choose two final concentrations of 100nM and 3 μM,

Second, the experimental method:

U20S cells stably expressing the EGFP-SlPi fusion protein, 5% CO _{2 were} cultured in DMEM high glucose medium containing 0.5 mg/ml G418 and 10% FBS. The cells were cultured in a 96-well black-bottomed cell culture plate according to the cells of 0.8x× ^{4 4} /Ι ΟΟμΙ/well, and cultured in 37 5% C0 _{2 for} 18-24 hours. The cells were washed once with 200 μl/well assay medium, and 150 μl/well assay medium was added and incubated for 37 min at 37 5% C0 ₂ . Formulation of agonist and working solutions of the present invention 5 - 21 and 23 - 24 compounds, adding 50 μl / well 4 agonist or 4 x compound or 4 x control working solution, 37 5% C0 _{2 was} incubated for 60 min, and the compound was repeated in parallel for 3 wells per concentration. Add 3 pre-warmed 3 fixatives ΙΟΟμΙ/well and mix at room temperature for 20 min. The cells were washed three times with 200 μl/well of staining solution and left in 200 μl/well staining solution for 1 h at room temperature. The cells were assayed on an IN Cel l Analyzer 1000 live cell imaging system under the following conditions: 20x objective, excitation wavelength Ex=460nm, emission wavelength Em=535nm, exposure to 300ms for detection of nuclear channel blue fluorescence; excitation wavelength Ex=475nm, The emission wavelength was Em=535 nm, and the cytoplasmic channel green fluorescent EGFP was detected at 800 ms exposure, and 5 fields of view per well were photographed continuously. Particle formation in the cytoplasm was analyzed using the GE IN Cel l Analyzer 1000 Mul ti Target Analys is Module software to calculate the activity of the compound stimulating S1Pi receptor endocytosis.

Compound agonistic intensity = (the average area of particles per cell in the compound treated group - the total area of particles per cell contained in the control treated group) / the total area of particles per cell contained in the control treated group.

2. 6 Data statistics and analysis:

The excitation intensity was calculated by taking the average of 15 fields of view (5 per hole) in 3 wells measured in parallel for each test concentration point of the compound.

Third, the experimental results

EGFP-S1P1-U20S cells are commercial cell lines used by the US Thermo Scientific to screen for compounds that have an effect on the SlPi receptor. After the SlPi receptor is activated, endocytosis is expressed as a cluster of green fluorescent protein reporter genes in cytoplasmic granules. Therefore, a compound having an agonistic activity against the S1P1 receptor can cause a change in the reporter molecule EGFP-S1P1 in this cell line.

Screening was performed using two final concentrations of 100 nM and 3 μ ,. The experimental results are shown in Table 1. Table 1

The compounds of Examples 15 and 23 of the present invention have significant agonistic activity.

Claims

Rights request

1. A compound of formula I or a geometric isomer or pharmaceutically acceptable salt or hydrate thereof:

among them-.

n is 1, 2 or 3,

X is 0, S, NH or Se,

Is hydrogen, an alkyl or acyl group having a carbon number of 1-8,

R ₂ is an alkyl or acyl substituent having a carbon number of 2 to 20, and the alkyl or acyl substituent may or may not contain an aromatic substituent, and each aromatic substituent may be substituted with 1 to 5 halogens, wherein the alkane a yl or acyl substituent which may contain from 1 to 3 sulfur, nitrogen or an oxygen atom to the alkyl group and the acyl group, an alkyl group and an aromatic substituent, an acyl group and an aromatic substituent, or a different or identical aromatic substituent, and

R ₃ is a hydrogen atom, an alkyl group having 1 to 8 carbon atoms, an acyl group or an alkoxy group.

2. A compound of formula I according to claim 1 or a geometric isomer or pharmaceutically acceptable salt or hydrate thereof, wherein said formula I is la:

among them-.

Is hydrogen, an alkyl or acyl group having a carbon number of 1-8, R ₂ is an alkyl or acyl substituent having a carbon number of 2 to 20, and the alkyl or acyl substituent may or may not contain an aromatic substituent, and each aromatic substituent may be substituted with 1 to 5 halogens, wherein the alkane a yl or acyl substituent which may contain from 1 to 3 sulfur, nitrogen or an oxygen atom to the alkyl group and the acyl group, an alkyl group and an aromatic substituent, an acyl group and an aromatic substituent, or a different or identical aromatic substituent, and

3. A compound of formula I according to claim 1 or a geometric isomer or pharmaceutically acceptable salt or hydrate thereof, wherein said Formula I is lb:

4. A compound of formula I according to claim 1 or a geometric isomer or pharmaceutically acceptable salt or hydrate thereof, wherein:

n is 1,

X is 0,

For hydrogen,

R ₂ is an alkyl or acyl substituent having a carbon number of 2 to 20, which is substituted by an alkyl group or an acyl group a group which may or may not contain an aromatic substituent, each of which may be substituted by 1 to 5 halogens, wherein the alkyl or acyl substituent may have 1 to 3 sulfur, nitrogen or an oxygen atom to the alkyl group and the acyl group. , an alkyl group and an aromatic substituent, an acyl group and an aromatic substituent, or a different or the same aromatic substituent, and

R ₃ is a hydrogen atom or a tert-butoxycarbonyl group.

5. A compound of formula I according to claim 1 or a geometric isomer or pharmaceutically acceptable salt or hydrate thereof, selected from the group consisting of:

(S)-2-amino-3-(4-octanoylphenyl)-propan-1-ol hydrochloride,

(S)-2-amino-3-(4-hexanoylphenyl)-propan-1-ol hydrochloride,

(S)-2-Amino-3-(4--heptanoylphenyl)-propan-1-ol hydrochloride,

(S)-2-amino-3-(4-(indolylphenyl)-propan-1-ol hydrochloride,

(S)-2-Amino-3-(4--n-undecylphenyl)-propan-1-ol hydrochloride,

(S)-2-Amino-3-(4--n-dodecanoylphenyl)-propan-1-ol hydrochloride,

(S)-2-Amino-3-(4-octylphenyl)-propan-1-ol hydrochloride,

(S)-2-amino-3-(4-hexylphenyl)-propan-1-ol hydrochloride,

(S)-2-amino-3-(4-heptylphenyl)-propan-1-ol hydrochloride,

(S)-2-Amino-3-(4--mercaptophenyl)-propan-1-ol hydrochloride,

(S)-2-Amino-3-(4--n-undecylphenyl)-propan-1-ol hydrochloride,

(S)-2-Amino-3-(4--n-dodecylphenyl)-propan-1-ol hydrochloride,

(S)-3-(4-benzyloxy-phenyl)-2-tert-butoxycarbonylamino-propan-1-ol,

(S)-3-(4-benzyloxy--phenyl)-2-amino-propan-1-ol trifluoroacetate,

(S)-3-[4-(3,4--fluoro-benzyloxy)-phenyl]-2-amino-propan-1-ol,

(S)-3-(4-(2,6-fluoro-benzyloxy)-phenyl]-2-amino-propan-1-ol, (S)-2-amino-3-(4-(4-bromo-phenoxy-acetyl)-phenyl-propan-1-ol, and

(S)-3-[4-(4-Methoxy-benzyloxy)-phenyl]-2-amino-propan-1-ol trifluoroacetate.

6. A pharmaceutical composition comprising at least one compound of the formula I according to any one of claims 1 to 5, or a geometric or pharmaceutically acceptable salt or hydrate thereof, and one or more pharmaceutically acceptable carriers Or an excipient.

7. A compound of formula I according to any one of claims 1 to 5, or a geometric isomer or pharmaceutically acceptable salt or hydrate thereof, or a pharmaceutical composition according to claim 6 for use in the preparation of a medicament for the treatment or prevention of rejection in organ transplantation and/or Or the use of drugs for autoimmune diseases.

8. The use of claim 7, wherein the autoimmune disease comprises, but is not limited to, multiple sclerosis, systemic lupus erythematosus, psoriasis or rheumatoid.

9. A method of treating or preventing rejection and/or autoimmune disease in organ transplantation, the method comprising administering to a patient at least one compound of the formula I according to any one of claims 1 to 5 or a geometric isomer thereof or a pharmaceutically acceptable drug thereof Use salt or hydrate.

10. The method of claim 9, wherein the autoimmune disease comprises, but is not limited to, multiple sclerosis, systemic lupus erythematosus, psoriasis or rheumatoid.