CN112694514A

CN112694514A - Pentacyclic triterpenoid TGR5 receptor agonist, and preparation method and application thereof

Info

Publication number: CN112694514A
Application number: CN201911011460.3A
Authority: CN
Inventors: 南发俊; 谢欣; 张晨露; 郭世猛; 卓宁; 梁晓影; 张仰明; 贠盈; 刘桦楠
Original assignee: Shanghai Institute of Materia Medica of CAS
Current assignee: Shanghai Institute of Materia Medica of CAS
Priority date: 2019-10-23
Filing date: 2019-10-23
Publication date: 2021-04-23
Anticipated expiration: 2039-10-23
Also published as: WO2021078150A1; CN112694514B

Abstract

The invention discloses a pentacyclic triterpenoid TGR5 receptor agonist, and a preparation method and application thereof. The pentacyclic triterpenoid TGR5 receptor of the inventionThe structure of the agonist is shown as a formula I, and the definition of each substituent is described in the specification and the claims. The pentacyclic triterpenoid compound has the advantages of increased solubility, improved permeability, remarkably improved TGR5 receptor agonistic activity, capability of penetrating Caco-2 monolayer cells and guarantee of in vivo drug effect exertion of the compound after oral administration. The TGR5 receptor agonist is expected to be further developed into a medicament for treating metabolic diseases represented by diabetes.

Description

Pentacyclic triterpenoid TGR5 receptor agonist, and preparation method and application thereof

Technical Field

The invention relates to TGR5 (bile acid G protein coupled receptor) agonists, in particular to pentacyclic triterpenoids, a preparation method thereof and application of the TGR5 receptor agonists or pharmaceutically acceptable salts thereof or a pharmaceutical composition containing any one of the TGR5 receptor agonists to preparation of medicines for treating diabetes, obesity, hyperlipidemia, liver injury and inflammatory diseases.

Background

TGR5 is a G protein-coupled receptor that can be activated by bile acids and is designated membrane bile acid receptor (M-BAR) or TGR 5. TGR5 is highly expressed in gallbladder, biliary epithelial cells, brown adipose tissue, muscle, gut, kidney, placenta, and brain. Binding of the ligand to TGR5 activates adenylate cyclase, which leads to elevated intracellular cAMP levels and different physiological effects in different tissues. In the small intestine, the elevation of cAMP level can promote the secretion of GLP-1, thus promoting the secretion of insulin and improving the sensitivity of the body to the insulin; in liver endothelial cells, it can promote the expression of endothelial NO synthetase and promote NO release, thereby protecting the endothelial cells from the damage of bile acid and lipid over-oxidation; in muscle and brown adipose tissue, the inactive thyroxine T4 can be converted to the active form T3 by activating cAMP-dependent deiodinase 2(DIO2), thereby increasing energy and oxygen consumption. Meanwhile, the activation of TGR5 has potential in treating inflammatory diseases such as atherosclerosis, colitis, nonalcoholic fatty liver disease and the like. Recent studies have shown that TGR5 activation can also promote mitochondrial fission and white adipose tissue remodeling. Therefore, the TGR5 receptor agonist is expected to be developed into a medicament for treating diabetes, obesity, hyperlipidemia, liver injury, inflammatory diseases and the like.

Currently under development TGR5 receptor agonists are largely classified into the following three types depending on the structural type: steroid TGR5 receptor agonists, triterpenoid TGR5 receptor agonists and synthetic small molecule TGR5 receptor agonists.

The synthetic micromolecule TGR5 receptor agonist has strong activity, but the compound can strongly activate gallbladder TGR5 receptors, so that smooth muscle is relaxed, and gallbladder filling is promoted, and meanwhile, the TGR5 receptor agonist can not only promote the release of neuropeptide in spinal cords in a central nervous system to cause pruritus, but also can promote the vasodilation of the whole body when the TGR5 receptors in the cardiovascular system are activated, so that the side effects of heart rate acceleration, cardiac rate increase and the like are caused, so the development of the intestinal tract selective TGR5 receptor agonist is a main strategy for avoiding toxic and side effects at present. Among them, Compound 12(J.Med.chem.,2018,61, 7589-7613) reported by Ardelyx and RDX-8940(Am J Physiol gateway Liver Physiol,2019,316, 412-761) of undisclosed structure preliminarily achieved evasion of toxicity of gall bladder for short-term administration by reducing molecular permeability.

Representative of steroidal TGR5 agonists INT777, currently in phase I clinical stage, has agonist activity on TGR5 receptors, EC₅₀Is 0.82 mu M, can obviously improve the energy consumption of obese C57BL/6 mice, relieve liver fatty lesion and increase the glucose tolerance and insulin sensitivity of the mice under the dosage of 30 mg/kg/d. However, the compound INT777 has the disadvantage of being difficult to prepare, starting from cholic acid, and being synthesizedThe route is up to twelve steps, the multi-step reaction conditions are extremely harsh, and recent literature also shows that INT777 has obvious gallbladder filling phenomenon (Finn, PD et al, Am J Physiol Gastrointest Liver Physiol,2019,316, 412-.

The pentacyclic triterpene natural product oleanolic acid can stimulate TGR5 receptors, and oral administration of 100mg/kg oleanolic acid can obviously reduce the body weight and blood sugar level of mice fed with high-fat diets, but due to the low activation activity of TGR5 receptors, the drug effect caused by off-target effect (PTP 1B receptor inhibition) cannot be excluded (biochem. Biophys. Res. Commun 2007,362,793-8). Betulinic acid, which also has a pentacyclic triterpene backbone, also has better TGR5 receptor agonistic activity (Journal of Medicinal chemistry.2008,51, 4849-4849; Journal of Medicinal Chemistry 2010,53,178-90), of which the compound 18dia 2 (EC) with the best activity₅₀0.12 μ M) (Journal of Medicinal Chemistry 2010,53,178-90) did not show significant weight and blood glucose lowering effects in mice raised on high fat, presumably due to poor physicochemical properties such as solubility and permeability and low bioavailability caused by the large lipid solubility of the betulinic acid parent nucleus.

The inventors have disclosed in WO2015135449 a class of pentacyclic triterpenoid TGR5 receptor agonists of the following structure:

wherein when R is₂Compound (C40, C99, C101, C103) human TGR5 agonistic activity (EC) when it is butyryloxy in alpha configuration₅₀237.8-455.7nM) with INT777 (EC)₅₀395.1 nM). The subsequent intensive study finds that the compound can obviously promote the secretion of GLP-1 from NCI-H716 cells in vitro, but the compound has extremely poor solubility due to high lipid solubility and cannot be dissolved in intestinal tracts, so the compound cannot take effect in vivo after oral administration. Caco-2 monolayer cell permeability experiment shows that the compound cannot permeate cell membranes, and TGR5 receptors are positioned on the basal membrane side of intestinal epidermal endocrine L cells, and insufficient permeability also determines that the compound cannot permeate intestinal epidermal inner partsThe L cell basement membrane is secreted to exert the drug effect.

Disclosure of Invention

The invention aims to provide a pentacyclic triterpenoid used as TGR5 receptor agonist.

In a first aspect of the invention, there is provided a compound of formula I, a pharmaceutically acceptable salt thereof, or a dimer thereof:

R₁is hydrogen, hydroxy, halogen or C1-C6 alkyl;

R₂is hydrogen, hydroxy, carboxy, halogen, C1-C6 alkoxy, 3-to 10-membered ring alkoxy, ═ O, ═ N-OH or R-Y (═ X)_m-O-; wherein m is 1 or 2; r is the following substituted or unsubstituted group: C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, 5-7 membered saturated heterocycle, R_aN (C1-C6 alkyl) -, R_aNH-or R_aO-; wherein each R is_aIndependently selected from the group consisting of substituted or unsubstituted: hydrogen, C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, or 5-7 membered saturated heterocyclyl; x is NH, O or S, Y is C, S or P;

R₃and R₄Each independently is unsubstituted or substituted C1-C6 alkyl; or R₃And C in position 3 to form an unsubstituted or substituted C3-C6 cycloalkyl group, R₄Is hydrogen, or unsubstituted or substituted C1-C6 alkyl;

R₅hydrogen, hydroxy, hydroxymethyl, formyl or carboxy;

R₆and R₇Each independently hydrogen, hydroxy, or C1-C6 alkyl;

R₈and R₉Each independently is hydrogen, hydroxy, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 3-6 membered oxygen containing heterocycle,

Wherein R is₁₂、R₁₃And R₁₄Is hydrogen, substituted or unsubstituted C1-C6 alkyl or R_c-M(＝R’)_r-; wherein r is 1 or 2; m is C, S or P, R' is O or S; r_cIs hydrogen, hydroxy, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 3-7 membered heterocyclyl, substituted or unsubstituted 4-8 membered heteroaryl, R_dNH-、R_dO(CH₂)_sNH-, or R_dO(CH₂)_s-; s is 0, 1, 2, 3 or 4, each R_dIndependently is hydrogen, substituted or unsubstituted C1-C6 alkyl, or substituted or unsubstituted 4-8 membered heteroaryl; a is O or S, B is NH or O, R₁₅Is hydrogen, substituted or unsubstituted C1-C6 alkyl, C3-C8 cycloalkyl, or a 5-7 membered saturated heterocyclic ring;

R₁₀is hydrogen, hydroxy, oxo (═ O), or C1-C6 alkyl;

R₁₁is hydrogen, hydroxy, oxo (═ O), or C1-C6 alkyl;

z is- (CH)₂) n-, n is 1, 2 or 3;

represents a single bond or a double bond;

each independently represents the R configuration, S configuration or racemic;

the substitution means that the hydrogen on the group is substituted by one or more substituents selected from the group consisting of: hydroxy, amino, methoxy, halogen, C1-C6 alkyl, C1-C6 alkoxy, oxo (═ O), epoxy, oxime (═ N-OH), carboxy (-COOH), sulfonic acid (-SO), and the like₂OH), glucosaminyl, morpholinyl, N-dimethyl, 4-methylpiperazinyl, pyridyl, piperidinyl, pyrrolidinyl.

In another preferred embodiment, R₁Is hydrogen, hydroxy or C1-C4 alkyl. In another preferred embodiment, R₁Is hydrogen.

In another preferred embodiment, R₂Is hydrogen, hydroxy, carboxy, halogen, C1-C4 alkoxy, 3-to 6-membered ring alkoxy, ═ O, ═ N-OH or R-Y (═ X)_m-O-；

Wherein m is 1 or 2;

r isSubstituted or unsubstituted of the following groups: C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, 5-7 membered saturated heterocycle, R_aN (C1-C4 alkyl) -, R_aNH-or R_aO-; wherein each R is_aIndependently substituted or unsubstituted: hydrogen, C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, or 5-7 membered saturated heterocyclyl;

x is NH, O or S;

y is C, S or P;

the substitution means that the hydrogen on the group is substituted by one or more substituents selected from the group consisting of: hydroxyl, methoxy, halogen, C1-C6 alkyl, C1-C6 alkoxy, carboxyl (-COOH), sulfonic acid (-SO)₂OH), glucosaminyl, morpholinyl, N-dimethyl.

In another preferred embodiment, for R₂The substitution means that the hydrogen on the group is substituted by one or more substituents selected from the group consisting of: hydroxyl, halogen, C1-C6 alkoxy, carboxyl (-COOH), sulfonic acid (-SO)₂OH), morpholinyl, N-dimethyl.

In another preferred embodiment, R₂Is carboxy, RCOO-, RCSO-or RSO₂O-, and R are as defined above. In another preferred embodiment, R is R_aNH-、R_aN (C1-C4 alkyl) -, R_aO, C1-C6 alkyl, C1-C6 alkoxy-C1-C6 alkylene or hydroxy-C1-C6 alkylene; wherein each R is_aIndependently hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, phenyl, halophenyl, C1-C4 alkoxy-substituted phenyl, imidazolyl, morpholinyl, carboxy-C1-C6 alkylene, sulfo-C1-C6 alkylene, hydroxy-substituted C1-C6 alkyl, morpholinyl-C1-C6 alkylene or N, N-dimethyl-C1-C6 alkylene.

In another preferred embodiment, R₂Attached to the ring in the alpha configuration.

In another preferred embodiment, R₃And R₄Each independently is a C1-C4 alkyl group;

or R₃And C at position 3 to form C1-C6 alkyl substituted C3-C6 cycloalkyl, R₄Is hydrogen.

In another preferred embodiment, R₃And R₄Each independently is a C1-C3 alkyl group. In another preferred embodiment, R₃And R₄And is also methyl.

In another preferred embodiment, R₅Hydroxymethyl, formyl or carboxyl. In another preferred embodiment, R₅Is a carboxyl group.

In another preferred embodiment, R₆And R₇Each independently hydrogen, methyl or ethyl.

In another preferred embodiment, R₈And R₉Each independently of the others is hydrogen, methyl, ethyl, n-propyl, isopropyl,

Wherein R is₁₂、R₁₃And R₁₄Each independently hydrogen, substituted or unsubstituted C1-C6 alkyl, R_cSO₂-, or R_cCO-wherein R_cIs hydrogen, hydroxyl, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 3-7 membered nitrogen-containing heterocyclic group (such as morpholine substituted piperidyl), R_dNH-、R_dO(CH₂)_sNH-、R_dO(CH₂)_s-; s is 0, 1, 2, 3 or 4; each R_dIs selected from hydrogen, substituted or unsubstituted C1-C6 alkyl, or substituted or unsubstituted 5-6 membered nitrogen containing heteroaryl (e.g., pyridyl);

b is NH or O, and B is NH or O,

R₁₅is hydrogen, substituted or unsubstituted C1-C6 alkyl, or 5-7 membered saturated heterocycle. Each substitution is as defined above.

In another preferred embodiment, R_cIs hydroxyl, morpholine substituted C1-C4 alkyl, carboxyl substituted C1-C4 alkyl, piperidine substituted C1-C4 alkyl,

In another preferred embodiment, R_dIs C1-C4 alkyl substituted by morpholine, C1-C4 alkyl substituted by pyrrolidine, C1-C4 alkyl substituted by piperidine, C1-C4 alkyl substituted by 4-methylpiperazine,Pyridine substituted C1-C4 alkyl, amino substituted C1-C4 alkyl, hydroxyl substituted C1-C6 alkyl, pyridyl, imidazolyl, amino substituted imidazolyl, and carboxyl substituted C1-C4 alkyl.

In another preferred embodiment, R₁₅Is C1-C4 alkyl substituted by carboxyl, morpholinyl, sulfo-C1-C6 alkylene, N-dimethyl-C1-C4 alkylene, morpholinyl-C1-C4 alkylene,

In another preferred embodiment, R₁₀Is hydrogen, hydroxy or oxo (═ O).

In another preferred embodiment, R₁₁Is hydrogen, hydroxy or oxo (═ O).

In another preferred embodiment, the compound is: any one of T1-T110.

The compounds of the present invention have asymmetric centers, chiral axes and chiral planes, and can exist in the form of racemates, R-isomers or S-isomers. The person skilled in the art is able to obtain the R-isomer and/or the S-isomer by resolution of the racemate by means of customary technical measures.

In a second aspect of the invention, there is provided a process for the preparation of a compound of the first aspect or a pharmaceutically acceptable salt or dimer thereof, said process being carried out in the presence of a pharmaceutically acceptable carrier

Introducing ester group, carbamate, sulfamate and carbonate into hydroxyl at the C3 position, diversifying double bond at the C20 position, introducing derivatizable groups such as hydroxyl and carboxyl, then performing ester formation, amide formation, carbamate, carbonate modification and the like, and finally removing a benzyl protecting group on the carboxyl at the C17 position by hydrogenation to obtain the compound of the first aspect, wherein the definition of each substituent is as described in the first aspect.

In a third aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of the first aspect, a pharmaceutically acceptable salt or dimer thereof; and

a pharmaceutically acceptable carrier or excipient.

The present invention provides a novel compound which can be used alone or in admixture with pharmaceutically acceptable adjuvants (e.g., excipients, diluents, etc.) to prepare tablets, capsules, granules, syrups, and the like for oral administration. The pharmaceutical composition can be prepared according to a conventional method in pharmacy.

In a fourth aspect of the invention there is provided a use of a compound of the first aspect or a pharmaceutical composition of the third aspect, (i) for the preparation of a bile acid G protein-coupled receptor (TGR5) agonist; or

(ii) Can be used for preparing medicine for treating metabolic diseases.

In another preferred embodiment, the metabolic disease is selected from the group consisting of: diabetes, obesity, hyperlipidemia, liver injury, and inflammatory diseases.

The present invention also provides a method of treating a metabolic disease by administering a compound of the first aspect or a pharmaceutical composition of the third aspect to a subject in need thereof.

The novel pentacyclic triterpenoid can effectively excite TGR5 receptors, can excite TGR5 receptors at nanomolar concentration, and has more excellent TGR5 exciting activity, more abundant natural sources and simpler synthetic method compared with the existing TGR5 agonists of some natural sources.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Not to be reiterated herein, but to the extent of space.

Detailed Description

The inventor of the present application has extensively and intensively studied, and a large polar group is introduced at an activity insensitive site of a pentacyclic triterpenoid (mainly, hydroxyl at position C3 is inverted, and ester bonds, carbamate, carbonate and other polar groups are introduced at positions C3 and C29), so that the physicochemical properties of the pentacyclic triterpenoid are remarkably improved, such as increased solubility and improved permeability, thereby overcoming the inherent defect of excessive fat solubility of the pentacyclic triterpenoid, and the TGR5 receptor agonistic activity is unexpectedly and remarkably improved, wherein the compound T75 humanized TGR5 agonistic activity is about 100 times of that of the compound C101(WO2015135449) in the previous invention, and the solubility of the compound is remarkably increased compared with the C101, and the compound can penetrate Caco-2 monolayer cells, so that the compound can be ensured to play the in-vivo efficacy after oral administration. Compared with INT777, the TGR5 receptor agonist not only has obviously improved physicochemical properties, but also has greatly improved TGR5 agonistic activity, and is expected to be further developed into a medicament for treating metabolic diseases represented by diabetes. On the basis of this, the present invention has been completed.

Term(s) for

In the present invention, the halogen is F, Cl, Br or I.

In the present invention, unless otherwise specified, the terms used have the ordinary meanings well known to those skilled in the art.

In the present invention, the term "C₁-C₆"means having 1, 2, 3, 4, 5 or 6 carbon atoms," C₁-C₈"means having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, and so forth. "3-10 membered" means having 3-10 ring atoms, and so on.

In the present invention, the term "alkyl" denotes a saturated linear or branched hydrocarbon moiety, for example the term "C₁-C₈Alkyl "refers to a straight or branched chain alkyl group having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, including, without limitation, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, and the like; preference is given to ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl.

In the present invention, the term "alkoxy" denotes-O-alkyl. For example, the term "C₁-C₆Alkoxy "means a straight or branched chain alkoxy group having 1 to 6 carbon atoms, including, but not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, and the like.

In the present invention, the term "alkenyl" denotes a straight or branched chain comprising at least one double bondHydrocarbyl moieties, e.g. the term "C₂-C₆The alkenyl group "means a straight or branched alkenyl group having 2 to 6 carbon atoms and containing one double bond, and includes, but is not limited to, ethenyl, propenyl, butenyl, isobutenyl, pentenyl, hexenyl, and the like.

In the present invention, the term "C₂-C₆Alkynyl "refers to a straight or branched chain alkynyl group having 2 to 6 carbon atoms containing one triple bond and includes, without limitation, ethynyl, propynyl, butynyl, isobutynyl, pentynyl, hexynyl, and the like.

In the context of the present invention, the term "cycloalkyl" denotes a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon moiety, for example the term "C₃-C₁₀Cycloalkyl "refers to a cyclic alkyl group having 3 to 10 carbon atoms in the ring, including, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, and the like. The term "C₃-C₈Cycloalkyl group "," C₃-C₇Cycloalkyl group ", and" C₃-C₆Cycloalkyl "has similar meaning. Polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl groups.

The term "cycloalkoxy" refers to-O- (cycloalkyl), wherein cycloalkyl is as defined above.

In the present invention, the term "heterocyclyl" denotes a cyclic group comprising at least one ring heteroatom (e.g. N, O or S). For example, the term "3-12 membered heterocyclic group" means a saturated or unsaturated 3-12 membered cyclic group containing 1 to 3 hetero atoms selected from oxygen, sulfur and nitrogen in the ring, such as dioxolanyl, tetrahydropyridinyl, dihydropyridinyl, dihydrofuranyl, dihydrothienyl, etc. The term "3-7 membered heterocyclyl" has a similar meaning.

In the present invention, the term "3-7 membered oxygen containing heterocyclic ring" means a cycloalkyl ring having 3-7 ring atoms and containing 1, 2 or 3O atoms, including without limitation a propylene oxide ring, a butylene oxide ring, a heptane oxide ring and the like.

In the present invention, the term "C₃-C₁₀Cycloalkenyl "means having from 3 to 10 carbons in the ringThe cyclic alkenyl group of atoms includes, but is not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclodecenyl and the like. The term "C₃-C₇Cycloalkenyl "has a similar meaning.

In the present invention, the term "aryl" denotes a hydrocarbon moiety comprising one or more aromatic rings, with a conjugated pi-electron system. For example, the term "C₆-C₁₂Aryl "refers to an aromatic ring group having 6 to 12 carbon atoms, such as phenyl, naphthyl, and the like, which does not contain heteroatoms in the ring. The term "C₆-C₁₀Aryl "has a similar meaning. Examples of aryl groups include, but are not limited to, phenyl (Ph), naphthyl, pyrenyl, anthracenyl, and phenanthrenyl.

In the present invention, the term "heteroaryl" denotes a moiety of an aromatic ring containing one or more heteroatoms (e.g., N, O or S), and examples of heteroaryl include furyl, fluorenyl, pyrrolyl, thienyl, oxazolyl, imidazolyl, thiazolyl, pyridyl, pyrrolinyl, pyrimidinyl, quinazolinyl, quinolinyl, pyranyl, isoquinolinyl, and indolyl.

The aryl ring may be fused to a heterocyclyl, heteroaryl or cycloalkyl ring, non-limiting examples of which include benzimidazole, benzothiazole, benzoxazole, benzisoxazole, benzopyrazole, quinoline, benzindole, chroman.

The heteroaryl group can be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring to which the parent structure is attached is a heteroaryl ring.

In the context of the present invention unless otherwise indicated,

indicates the attachment site.

Unless otherwise specified, alkyl, alkoxy, cycloalkyl, heterocyclyl and aryl groups described herein are substituted and unsubstituted groups. Possible substituents on the alkyl, alkoxy, cycloalkyl, heterocyclyl and aryl groups include, but are not limited to: hydroxyl, amino, nitro, nitrile, halogen, C1-C6 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C20 cycloalkyl, C3-C20 cycloalkenyl, C1-C20 heterocycloalkyl, C1-C20 heterocycloalkenyl, C1-C6 alkoxy, aryl, heteroaryl, heteroaryloxy, C1-C10 alkylamino, C1-C20 dialkylamino, arylamino, diarylamino, C1-C10 alkylsulfamoyl, arylsulfamoyl, C1-C10 alkylimino, C1-C10 alkylsulfamomino, arylsulfonylimino, mercapto, C1-C10 alkylthio, C1-C10 alkylsulfonyl, arylsulfonyl, acylamino, aminoacyl, aminothioacyl, guanidino, ureido, acyl, thioacyl, acyloxy, carboxyl, and carboxylate. In another aspect, cycloalkyl, heterocycloalkyl, heterocycloalkenyl, aryl, and heteroaryl groups can also be fused to each other.

In the invention, the substitution is mono-substitution or multi-substitution, and the multi-substitution is di-substitution, tri-substitution, tetra-substitution or penta-substitution. By disubstituted is meant having two substituents and so on.

Unless otherwise specified, the structural formulae depicted herein are intended to include all tautomeric, enantiomeric and stereoisomeric forms (e.g., enantiomers, diastereomers, geometric isomers or conformational isomers): for example, the R, S configuration containing an asymmetric center, the (Z), (E) isomers and the conformational isomers of (Z), (E) of the double bond. Thus, individual stereochemical isomers, tautomers or enantiomers, diastereomers or geometric isomers or conformational isomers or mixtures of tautomers of the compounds of the present invention are within the scope of the present invention.

The term "tautomer" means that structural isomers having different energies may exceed the low energy barrier and thus be converted to each other. For example, proton tautomers (i.e., proton shift) include interconversion by proton shift, such as 1H-indazole and 2H-indazole, 1H-benzo [ d ] imidazole and 3H-benzo [ d ] imidazole, and valence tautomers include interconversion by some bond-forming electron recombination.

Herein, the pharmaceutically acceptable salt is not particularly limited, and preferably includes: inorganic acid salts, organic acid salts, alkylsulfonic acid salts and arylsulfonic acid salts; the inorganic acid salt comprises hydrochloride, hydrobromide, nitrate, sulfate, phosphate and the like; the organic acid salt comprises formate, acetate, propionate, benzoate, maleate, fumarate, succinate, tartrate, citrate and the like; the alkyl sulfonate includes methyl sulfonate, ethyl sulfonate and the like; the aryl sulfonate includes benzene sulfonate, p-toluene sulfonate and the like.

Preparation method

The pentacyclic triterpenoid can be prepared by the following route.

Route 1

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Pharmaceutical composition

The invention also provides a pharmaceutical composition comprising a safe and effective amount of the active ingredient, and a pharmaceutically acceptable carrier.

The active ingredient refers to the compound of the formula I.

The active ingredient and the pharmaceutical composition are used for preparing the medicines for treating the metabolic diseases. The "active ingredient" and pharmaceutical compositions described herein are useful as TGR5 receptor agonists. In another preferred embodiment, for the preparation of a medicament for the prevention and/or treatment of diseases modulated by TGR5 agonists.

"safe and effective amount" means: the amount of active ingredient is sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000mg of active ingredient per dose, more preferably, 10-200mg of active ingredient per dose. Preferably, said "dose" is a tablet.

"pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. By "compatible" is meant herein that the components of the composition are capable of being combined with the active ingredients of the present invention and with each other without significantly diminishing the efficacy of the active ingredient. Examples of pharmaceutically acceptable carrier moieties are cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), and the like

) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

The mode of administration of the active ingredient or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and the like.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, especially cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like. In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active ingredients, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these materials, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

The compounds of the present invention may be administered alone or in combination with other therapeutic agents.

When the pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is suitable for mammals (such as human beings) to be treated, wherein the administration dose is a pharmaceutically-considered effective administration dose, and for a human body with a weight of 60kg, the daily administration dose is usually 1 to 2000mg, preferably 20 to 500 mg. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures for which specific conditions are not indicated in the following examples are generally carried out according to conventional conditions (e.g.as described in Sambrook et al, molecular cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989)) or according to the conditions as recommended by the manufacturer. Unless otherwise indicated, percentages and parts are percentages and parts by weight.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

In the following preparation examples, NMR was measured with a Mercury-Vx 300M instrument manufactured by Varian, and NMR was calibrated: Δ H7.26 ppm (CDCl)₃)，2.50ppm(DMSO-d₆) (ii) a Measuring the mass spectrum by an Agilent 1200 Quadrupole LC/MS liquid chromatograph-mass spectrometer or SHIMADZU GCMS-QP 5050A; reagents are mainly provided by Shanghai chemical reagents company; TLC thin layer chromatography silica gel plate is produced by Shandong tobacco Taihuyou silica gel development Co., Ltd, model number HSGF 254; the normal phase column chromatography silica gel used for compound purification is produced by Shandong Qingdao ocean chemical plant, model zcx-11, 200-300 mesh.

The abbreviations herein correspond to the following Chinese: DMAP 4-dimethylaminopyridine; DCM is dichloromethane; DMF: n, N-dimethylformamide; THF: tetrahydrofuran.

Example 1

(1) The raw material betulinic acid S1(1.2g,2.63mmol) was dissolved in methanol (50mL) at room temperature, after nitrogen exchange, 10% Pd/C was added quickly, after nitrogen exchange, hydrogen exchange was performed, and stirring was performed at room temperature. The reaction was complete after 24 hours by TLC. After nitrogen exchange, Pd/C is filtered out, after reaction liquid is dried in a spinning mode, column chromatography separation is carried out by using an eluent system with petroleum ether/ethyl acetate as 10:1, and a compound S21.04g (2.27mmol) is obtained as a white solid, and the yield is as follows: 86 percent.¹H NMR(300MHz,CDCl₃)δ3.13(t,1H,J＝9.0,6.9Hz),2.28-2.16(m,2H),1.98-1.78(m,4H),1.64-0.96(m, other alicyclic protons), 0.96(s,3H),0.93(s,3H),0.92(s,3H),0.90(s,3H),0.89(s,3H),0.87(s,3H),0.78(s,3H).

(2) The product S2 from the above step (1.04g,2.27mmol) was dissolved in DMF (20mL) at room temperature, anhydrous potassium carbonate (0.626g,4.54mmol) was added, and benzyl chloride (0.313mL,2.72mmol) was slowly added dropwise with stirring. After the completion of the dropwise addition, the reaction solution was transferred to 50 ℃ and stirred for 3 hours, and the completion of the reaction was monitored by TLC. The mixture was cooled to room temperature, diluted with 50mL of deionized water, extracted with ethyl acetate (2 × 50mL), and the combined organic layers were washed with deionized water and saturated brine, respectively, dried over sodium sulfate and distilled under reduced pressure to give the desired compound s31.21g (2.20mmol) as a white solid in the next reaction in yield: 97 percent.

(3) S3(1.21g, 2.20mmol) was dissolved in dichloromethane (30mL) under an ice-water bath, Dess-Martin oxidant (1.87g,4.40mmol) was added in portions, allowed to warm slowly to room temperature and stirred for 1 hour. The reaction mixture was filtered, spin dried and separated by column chromatography using an eluent system of petroleum ether/ethyl acetate 10:1 to give compound s40.983g (1.80mmol) as a white solid in yield: 82 percent.¹H NMR(300MHz,CDCl₃) δ 7.38-7.30(m,5H),5.16-5.05(m,2H),2.55-2.34(m,2H), 2.31-2.18 (m,3H), 1.95-1.12 (m, other alicyclic protons), 1.06(s,3H),1.01(s,3H),0.93(s,3H),0.91(s,3H),0.86(d,3H, J ═ 6.9Hz),0.79-0.73(m,6H).

(4) Product S4 from the previous step (0.983g,1.80mmol) and S- (-) -2-methyloxazaborolidine (100mg,0.36mmol) were taken in a 100mL oven dried round bottom flask and fresh sodium treated THF (70mL) was added. 10M borane-tetrahydrofuran solution (0.32mL) was slowly added dropwise at room temperature over ten minutes with controlled rate, stirred at room temperature for ten minutes and TLC monitored to show that the reaction had progressed. Moving the reaction bottle to an ice-water bath, slowly dropwise adding methanol to quench the reaction, spin-drying the solvent after no more bubbles are generated, and performing column chromatography separation by using an eluent system with petroleum ether/ethyl acetate as a 10:1 to obtain a compound S58 5827 mg (1.51mmol) which is a white solid and has the yield: 84 percent.¹H NMR(300MHz,CDCl₃)δ7.37-7.30(m,5H),5.15-5.06(m,2H),3.38(t,1H,J＝3.0Hz),2.55-2.34(m,2H) 2.31-2.18 (m,3H), 1.95-1.12 (m, other alicyclic protons), 0.94(s,3H),0.93(s,3H),0.91(s,3H),0.85-0.82(m,9H),0.75-0.73(m,6H).

(5) Intermediate S5(100mg,0.182mmol) was dissolved in dry DCM (5mL), trichloroacetyl isocyanate (25.9. mu.L, 0.218mmol) was added dropwise at 0 deg.C, the temperature was maintained and the reaction was allowed to proceed for 1 hour, DCM was spun off, methanol (5mL) was added and dissolved, then potassium carbonate (50.2mg,0.364mmol) was added at room temperature and stirred for 2 hours, TLC showed completion of the reaction. Quenching with deionized water, removing methanol by centrifugation, extracting with ethyl acetate (3 × 30mL), washing the combined organic layers with deionized water and saturated brine, drying over sodium sulfate, concentrating, and separating by column chromatography to obtain product S6103 mg (0.174mmol), yield: 95.5 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.29(m,5H),5.15-5.06(m,2H),4.59(br s,2H),4.49(t,1H, J ═ 3.0Hz),2.30-2.15(m,3H),1.92-0.99(m, other alicyclic protons), 0.97(s,3H),0.87-0.83(m,12H),0.75-0.73(m,6H).

(6) Preparation of compound T1: intermediate S6(103mg,0.174mmol) was dissolved in methanol (20mL) at room temperature, after purging with nitrogen, 10% Pd/C was added quickly, after purging with nitrogen, with hydrogen, and stirred at room temperature. The reaction was complete after 2 hours by TLC. After nitrogen exchange, Pd/C is filtered out, after reaction liquid is dried in a spinning mode, column chromatography separation is carried out by using an eluent system of petroleum ether/ethyl acetate, and a compound T179.0 mg is obtained and is a white solid, and the yield is as follows: 90.5 percent.¹H NMR(500MHz,CDCl₃) δ 4.86(br s,2H),4.50(t,1H, J ═ 3.0Hz),2.28-2.21(m,3H),1.90-1.09(m, other alicyclic protons), 1.00(s,3H),0.94(s,3H),0.88-0.85(m,12H),0.76(d,3H, J ═ 6.5Hz).

Example 2

(1) S5(1.00g,1.82mmol) was dissolved in dry DCM (10mL), pyridine (732. mu.L, 9.10mmol) was added dropwise at 0 deg.C, after 0.5 hr p-nitrophenyl chloroformate (1.10g,5.46mmol) in dry DCM (10mL) was added dropwise, and after returning to room temperature, the reaction was stirred for 5 hr, as indicated by TLC. DCM was spun off, extracted with ethyl acetate (3X 100mL), and the mixtures were combinedThe organic layer was washed with deionized water and saturated brine, dried over sodium sulfate, concentrated, and separated by column chromatography to give S71.20g (1.69mmol), yield: 92.2 percent.¹H NMR(300MHz,CDCl₃) δ 8.30-8.27(m,2H),7.40-7.35(m,7H),5.15-5.06(m,2H),4.60(t,1H, J ═ 3.0Hz),2.30-2.15(m,5H),2.04-1.05(m, other alicyclic protons), 0.96(s,3H),0.95(s,3H),0.92(s,3H),0.86-0.84(m,6H),0.76-0.74(m,6H).

(2) Intermediate S-7(100mg,0.182mmol), DMAP (66.7mg,0.546mmol) and triethylamine (75.7. mu.L, 0.546mmol) were dissolved in dry DCM (2mL), cyclopropylamine (63.0. mu.L, 0.910mmol) was added dropwise, stirring overnight at room temperature, the next day TLC showed complete reaction. The DCM was spun off, extracted with ethyl acetate (3 × 50mL), the combined organic layers were washed with deionized water and saturated brine respectively, dried over sodium sulfate, concentrated and isolated by column chromatography to give 81mg (0.128mmol) of the product.¹H NMR(300MHz,CDCl₃) δ 7.38-7.30(m,5H),5.15-5.06(m,2H),4.82(br s,1H),4.51(t,1H, J ═ 2.7Hz),2.61-2.57(m,1H),2.30-2.15(m,3H),1.92-0.98(m, other alicyclic protons), 0.95(s,3H),0.86-0.83(m,12H),0.75-0.68(m,8H),0.55-0.50(m,2H).

(3) The compound T653.5mg can be obtained by a similar debenzylation process, and the yield is 77.0%.¹H NMR(300MHz,CDCl₃) δ 4.51(t,1H, J ═ 3.0Hz),2.60-2.55(m,1H),2.29-2.17(m,3H),1.91-1.07(m, other alicyclic protons), 0.97(s,3H),0.93(s,3H),0.87-0.84(m,12H),0.76-0.68(m,5H),0.56-0.51(m,2H).

Using a similar procedure as in example 2, intermediate S7 was reacted with a different amine to give the following compounds.

Example 3

(1) Preparation of intermediate S8: sodium hydride (9.84mg,0.246mmol, 60%) was suspended in dry DMF (1mL), 2mL of a dry DMF solution of intermediate S8(76.0mg, 0.123mmol) was added dropwise at 0 deg.C, methyl iodide (38.3. mu.L, 0.615mmol) was added dropwise while maintaining at 0 deg.C, and after 10 minutes, the mixture was returned to room temperature and stirred overnight. The next day TLC monitoring indicated that the reaction was complete. Quenched by addition of deionized water, extracted with ethyl acetate (3 × 50mL), and the combined organic layers were washed with deionized water and saturated brine, respectively, dried over sodium sulfate, concentrated, and separated by column chromatography to give the product, s 959.0 mg (0.093mmol), yield: 76.0 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.30(m,5H),5.15-5.06(m,2H),4.49(t,1H, J ═ 2.7Hz),3.35-3.28(m,2H),2.90(s,3H),2.30-2.15(m,3H),1.91-0.96(m, other alicyclic protons), 0.93(s,3H),0.88-0.84(m,12H),0.76-0.73(m,6H).

(2) Intermediate S9 was subjected to a similar debenzylation procedure to give compound T20(32.0mg, 63.2%).¹H NMR(300MHz,CDCl₃) δ 4.50(t,1H, J ═ 2.7Hz),3.36-3.29(m,2H),2.91(s,3H),2.29-2.17(m,3H),1.92-1.08(m, other alicyclic protons), 0.97(s,3H),0.94(s,3H),0.88-0.85(m,12H),0.76(d,3H, J ═ 6.9Hz).

Example 4

(1) Preparation of intermediate S10: compound S5(130mg,0.237mmol) was dissolved in 3mL pyridine, ethyl chloroformate (113. mu.L, 1.19mmol) was added at-20 deg.C, and after 10 min stirring was returned to room temperature and after 2h the reaction was complete by TLC. Diluting with 30mL of deionized water, extracting with ethyl acetate (3X 50mL), washing the combined organic layers with dilute hydrochloric acid, deionized water and saturated brine respectively, drying over sodium sulfate, concentrating, and separating by column chromatography to obtain S1097.0 mg (0.156mmol), yield: 66.0 percent.¹H NMR(300MHz,CDCl₃) δ 7.38-7.30(m,5H),5.15-5.06(m,2H),4.46(t,1H, J ═ 3.0Hz),4.19(q,2H, J ═ 7.2Hz),2.30-2.14(m,3H),1.97-0.98(m, other alicyclic protons), 0.95(s,3H),0.90-0.85(m,9H),0.83(s,3H),0.75-0.73(m,6H).

(2) Preparation of compound T21: intermediate S10(97.0mg,0.156mmol) was dissolved in ethyl acetate and t-butanol (1:1,10mL) and after purging, 10% Pd (OH) was added quickly₂and/C, changing nitrogen and hydrogen, and stirring at room temperature. The reaction was complete after 1 hour by TLC. After nitrogen exchange, Pd (OH) is filtered out₂And C, after the reaction solution is dried in a spinning mode, performing column chromatography separation to obtain a compound T2148.2mg (0.091mmol), wherein the yield is as follows: 58.1 percent.¹H NMR(300MHz,CDCl₃) δ 4.47(t,1H, J ═ 3.0Hz),4.20(q,2H, J ═ 7.2Hz),2.28-2.14(m,3H),1.96-1.14(m, other alicyclic protons), 0.98(s,3H),0.93(s,3H),0.90(s,3H),0.88-0.84(m,9H),0.75(d,3H, J ═ 6.9Hz).

Example 5

(1) Intermediate S5(50.0mg, 0.091mmol) was dissolved in 1mL pyridine solution, followed by dropwise addition of methylaminosulfonyl chloride (39.3. mu.L, 0.455mmol) while maintaining 0 deg.C, after 10 minutes returning to room temperature, stirring overnight, and TLC the next day indicated complete reaction. The pyridine was spun off, deionized water was added, extraction was performed with ethyl acetate (3 × 30mL), the combined organic layers were washed with deionized water and saturated brine, respectively, dried over sodium sulfate, concentrated, and separated by column chromatography to give 51.0mg (0.080mmol) of the product, yield: 87.2 percent.¹H NMR(300MHz,CDCl₃) δ 7.38-7.30(m,5H),5.15-5.06(m,2H),4.33-4.28(m,2H),2.82(d,3H, J ═ 5.1Hz),2.30-2.15(m,3H),1.95-1.10(m, other alicyclic protons), 1.00(s,3H),0.95(s,3H),0.88(s,3H),0.86-0.83(m,6H),0.74(t,6H, J ═ 3.3Hz).

(2) Compound T22(28.0mg, 90.5%) was obtained via a debenzylation procedure analogous to that used in the preparation of compound T1.¹H NMR(300MHz,CDCl₃) δ 4.40-4.33(m,1H),4.30(t,1H, J ═ 2.7Hz),2.83(d,3H, J ═ 5.4Hz),2.28-2.16(m,4H),1.96-1.14(m, other alicyclic protons), 1.01(s,3H),0.99(s,3H),0.93(s,3H),0.89-0.84(m,9H),0.76(d,3H,J＝6.6Hz).

Using a similar procedure as in example 5, intermediate S5 was reacted with acid chloride, sulfonyl chloride and aminosulfonyl chloride to provide compounds T23-T26.

Example 6

(1) Intermediate S5 was subjected to a similar debenzylation procedure to give compound S11(35.0mg, 87.5%).¹H NMR(300MHz,CDCl₃) δ 3.39(s,1H),2.28-2.16(m,2H),1.98-1.78(m,4H),1.64-0.96(m, other alicyclic protons), 0.96(s,3H),0.93(s,3H),0.92(s,3H),0.90(s,3H),0.89(s,3H),0.87(s,3H),0.78(s,3H).

(2) Sodium hydride (15.3mg,0.382mmol) was suspended in 1mL of dry THF, stirred at 0 deg.C for 0.5h, then ethylthioisocyanate (100. mu.L, 3.82mmol) was added and stirred overnight back to room temperature, the next day TLC indicated complete reaction. After quenching with deionized water, THF was spun off, extracted with ethyl acetate (3 × 30mL), and the combined organic layers were washed with deionized water and saturated brine, respectively, dried over sodium sulfate, concentrated, and separated by column chromatography to give the product, tj2726.0 mg (0.048mmol), yield: 62.4 percent.¹H NMR(400MHz,CDCl₃) δ 6.71(t,0.5H, J ═ 5.6Hz),6.24(t,0.5H, J ═ 5.6Hz),5.20(t,1H, J ═ 2.8Hz),3.64 to 3.57(m,1H),3.37 to 3.31(m,1H),2.25 to 2.20(m,4H),1.91 to 1.05(m, other fat ring protons), 0.97 to 0.91(m,9H),0.89 to 0.82(m,12H),0.75(d,3H, J ═ 6.4Hz).

Example 7

(1) After protection of the benzyl betulinate, the hydroxyl group is inverted to obtain an intermediate S12.¹H NMR(300MHz,CDCl₃) δ 7.37-7.31(m,5H),5.17-5.06(m,2H),4.72(s,1H),4.60-4.58(m,1H),3.39-3.36(m,1H),3.06-2.98(m,1H),2.29-2.13(m,3H),1.98-1.02(m, other alicyclic protons), 0.96(s,3H),0.92(s,3H),0.81(s,6H),0.76(s,3H).

(2) S12(1.96g,3.59mmol) and DMAP (43.9mg,0.359mmol) were dissolved in pyridine (20mL), acetic anhydride (1.70mL,18.0mmol) was added dropwise at 0 deg.C, after 10 minutes to room temperature and stirred overnight, the next day TLC monitoring indicated that the reaction was complete. The pyridine was spun off, extracted with ethyl acetate (3 × 100mL), the combined organic layers were washed with deionized water, 1N dilute hydrochloric acid and saturated brine, respectively, dried over sodium sulfate, concentrated, and isolated by column chromatography to give product s132.07g (3.52mmol), yield: 98.1 percent.¹H NMR(300MHz,CDCl₃) δ 7.39-7.31(m,5H),5.17-5.07(m,2H),4.72(s,1H),4.62-4.58(m,2H),3.07-2.98(m,1H),2.31-2.14(m,3H),2.06(s,3H),1.94-1.03(m, other fat ring protons), 1.68(s,3H),1.00(s,3H),0.86(s,3H),0.82(s,6H),0.77(s,3H).

(3) Intermediate S13(60.0mg,0.102mmol) was dissolved in MeOH/DCM (1:1,10mL), ozone was bubbled at-78 deg.C, and TLC after 1 min indicated completion of the reaction. After the introduction of ozone was stopped, residual ozone was exhausted with oxygen, and 25. mu.L of dimethyl sulfide was added to quench the reaction, and the reaction was warmed to room temperature and stirred overnight. The next day, the reaction solution was spin-dried and then separated by column chromatography to give compound s1444.0mg (0.075mmol), yield: 73.1 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.31(m,5H),5.17-5.07(m,2H),4.60(t,1H, J ═ 3.0Hz),3.29-3.22(m,1H),2.33-2.27(m,1H),2.16(s,3H),2.07(s,3H),2.04-1.06(m, other alicyclic protons), 1.02(s,3H),0.85(s,3H),0.81(s,6H),0.74(s,3H).

(4) Intermediate S14 was subjected to a similar debenzylation procedure to give compound T28(35.5mg, 95.2%).¹H NMR(300MHz,CDCl₃) δ 4.61(s,1H),3.29-3.20(m,1H),2.31-2.25(m,1H),2.18(s,3H),2.08(s,3H),2.04-1.09(m, other alicyclic protons), 1.06(s,3H),0.93(s,3H),0.86(s,3H),0.84(s,3H),0.83(s,3H).

Example 8

(1) Intermediate S13(70.0mg,0.119mmol) was dissolved in DCM (3mL), m-CPBA (51.3mg,0.238mmol, 80%) was added at 0 deg.C, and after 10 min returned to room temperature and stirred for 2h, TLC monitoring indicated the reaction was complete. Quenching with saturated aqueous sodium sulfite solution, extracting DCM and then with ethyl acetate (3 × 30mL), washing the combined organic layers with deionized water, saturated aqueous sodium bicarbonate solution and saturated brine respectively, drying over sodium sulfate and concentrating, separating by column chromatography to obtain product s1558.0 mg (0.096mmol), yield: 80.7 percent.¹H NMR(300MHz,CDCl₃) δ 7.35-7.32(m,5H),5.11-5.08(m,2H),4.61(t,1H, J ═ 3.0Hz),2.65-2.54(m,2H),2.32-2.27(m,1H),2.17-2.10(m,2H),2.07(s,3H),1.97-1.02(m, other alicyclic protons), 1.23(s,3H),0.99(s,3H),0.86(s,3H),0.82(s,6H),0.76(s,3H).

(2) Intermediate S15 was subjected to a similar debenzylation procedure to give compound T29(27.2mg, 55.1%).¹H NMR(300MHz,CDCl₃) δ 4.62(t,1H, J ═ 2.4Hz),2.66-2.63(m,2H),2.30-2.27(m,1H),2.18-2.11(m,2H),2.08(s,3H),2.04-1.05(m, other alicyclic protons), 1.25(s,3H),1.02(s,3H),0.94(s,3H),0.87(s,3H),0.86(s,3H),0.84(s,3H).

Example 9

(1) Intermediate S14(25.0mg,0.043mmol) was dissolved in pyridine (1mL), hydroxylamine hydrochloride (34.7mg,0.500mmol) was added at 0 ℃, after 10 min to room temperature and stirred overnight, the next day TLC showed complete reaction. Most of the pyridine was spun off, extracted with ethyl acetate (3 × 30mL), and the combined organic layers were washed with deionized water, 1N diluted hydrochloric acid and saturated brine, respectively, dried over sodium sulfate, concentrated, and separated by column chromatography to give the product, s 1616.0 mg (0.026mmol), yield: 62.2 percent.¹H NMR(300MHz,CDCl₃)δ7.38-7.30(m,5H),5.17-5.08(m,2H),4.61(t,1H,J2.7Hz),3.21-3.13(m,1H),2.33-2.28(m,1H),2.21-2.11(m,2H),2.06(s,3H),2.04-1.00(m, other alicyclic protons), 1.81(s,3H),0.98(s,3H),0.86(s,3H),0.82(s,6H),0.75(s,3H).

(2) Intermediate S16 was subjected to a similar debenzylation procedure to give compound T30(6.00mg, 44.1%).¹H NMR(500MHz,CDCl₃) δ 4.60(t,1H, J ═ 3.0Hz),3.16(t,1H, J ═ 11.0Hz),2.38-2.36(m,1H),2.24-2.20(m,1H),2.11-2.03(m,5H),1.90(s,3H),1.86-1.75(m,3H),1.61-1.03(m, other fat ring protons), 1.00(s,3H),0.92(s,3H),0.84(s,3H),0.82(s,3H),0.81(s,3H).

Example 10

(1) S13(53.0mg,0.090mmol) was dissolved in dry THF (1mL) and 10M BH was added dropwise at 0 deg.C₃-Me₂S (27.0. mu.L, 0.270mmol), stirred for 1 hour and then checked by TLC for substantial completion of the reaction. Ethanol (100. mu.L), saturated sodium carbonate solution (80. mu.L) and 30% hydrogen peroxide solution (100. mu.L) were added to the reaction solution in this order, and the mixture was stirred at room temperature for 2 hours to complete the reaction as detected by TLC. The remaining hydrogen peroxide was neutralized by adding saturated sodium sulfite solution, extracted with ethyl acetate (3 × 30mL), and the combined organic layers were washed with deionized water, 1N dilute hydrochloric acid and saturated brine, respectively, dried over sodium sulfate and concentrated, and subjected to column chromatography to give compound s 1739.0 mg (0.065mmol), yield: 71.6 percent.¹H NMR(300MHz,CDCl₃) δ 7.36-7.30(m,5H),5.15-5.05(m,2H),4.61(t,1H, J ═ 3.0Hz),3.77(dd,1H, J ═ 10.5Hz,4.8Hz),3.41(dd,1H, J ═ 10.5Hz,7.8Hz),2.35-2.16(m,3H),2.07(s,3H),1.90-1.80(m,3H),1.69-1.03(m, other alicyclic protons), 0.98(s,3H),0.95(d,3H, J ═ 6.9Hz),0.86(s,3H),0.82(s,6H),0.75(s,3H).

(2) Intermediate S17 was subjected to a similar debenzylation procedure to give compound T31(8.00mg, 55.3%).¹H NMR(500MHz,CDCl₃) δ 4.62(t,1H, J ═ 3.0Hz),3.79(dd,1H, J ═ 10.5,4.5Hz),3.44(dd,1H, J ═ 10.5,8.0Hz),2.35-2.30(m,1H),2.28-2.19(m,2H),2.08(s,3H),1.91-1.84(m,3H),1.75-1.08(m, other fat ring protons), 1.01(s,3H),0.97 (J ═ 3.0Hz),3.79(dd,1H, J ═ 10.5 Hz),2.35-2.30(m,1H),2.28-2.19(m,2H),2d,3H,J＝6.5Hz),0.94(s,3H),0.87(s,3H),0.86(s,3H),0.83(s,3H).

Example 11

(1) Will be SeO₂(11.3mg,0.102mmol) and TBHP (39.1. mu.L, 0.408mmol) were dissolved in dry DCM (1mL), acetic acid (1.14. mu.L, 0.020mmol) was added dropwise at 0 deg.C, after stirring for 10 minutes, a solution of intermediate S13(120mg,0.204mmol) in dry DCM (3mL) was added and stirred overnight at room temperature, and the next day TLC checked for reaction completion. The remaining TBHP was neutralized by addition of saturated sodium sulfite solution, DCM was decanted, ethyl acetate (3X 30mL) was used for extraction, and the combined organic layers were washed with deionized water and saturated Na, respectively₂SO₃The solution is washed with saturated brine, dried over sodium sulfate and concentrated, then dissolved in, for example, methanol (5mL), and NaBH is added₄(15.4mg,0.408mmol) for 1 hour and TLC check reaction complete. Adding acetone for quenching, directly carrying out rotary drying, and then carrying out column chromatography separation to obtain a compound S1885.4mg (0.141mmol), wherein the yield is as follows: 69.3 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.32(m,5H),5.17-5.07(m,2H),4.95(s,1H),4.91(s,1H),4.61(t,1H, J ═ 3.0Hz),4.13-4.10(m,2H),2.93-2.85(m,1H),2.34-2.28(m,1H),2.21-2.12(m,2H),2.06(s,3H),1.98-1.03(m, other alicyclic protons), 1.00(s,3H),0.86(s,3H),0.82(s,6H),0.76(s,3H).

(2) Intermediate S18(45.0mg,0.075mmol) was dissolved in MeOH/DCM (1:1,10mL), and ozone was bubbled through at-78 deg.C, after 1 min TLC indicated complete reaction. After the introduction of ozone was stopped, residual ozone was exhausted with oxygen, and 25. mu.L of dimethyl sulfide was added to quench the reaction, and the reaction was warmed to room temperature and stirred overnight. The next day, the reaction solution was spin-dried and subjected to silica gel column chromatography to obtain compound S1931.0 mg (0.051mmol), yield: 68.7 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.32(m,5H),5.18-5.07(m,2H),4.61(t,1H, J ═ 3.0Hz),4.27(d,2H, J ═ 4.5Hz),3.28-3.19(m,1H),3.11(t,1H, J ═ 4.8Hz),2.35-2.20(m,2H),2.07(s,3H),2.04-1.05(m, other aliphatic ring protons), 1.01(s,3H),0.86(s,3H),0.81(s,6H),0.74(s,3H).

(3) Preparation of compound T32: intermediate S19 was similarly preparedThe debenzylation procedure of (2) gave compound T32(21.1mg, 79.9%).¹H NMR(300MHz,CDCl₃) δ 4.62(s,1H),4.30(s,2H),3.27-3.18(m,2H),2.33-2.25(m,2H),2.08(s,3H),2.05-1.08(m, other alicyclic protons), 1.05(s,3H),0.92(s,3H),0.86(s,3H),0.84(s,3H),0.83(s,3H).

Example 12

(1) S18(40.4mg,0.067mmol) was dissolved in dry THF (1mL) and 10M BH was added dropwise at 0 deg.C₃-Me₂S (20.1. mu.L, 0.201mmol), stirred for 1 hour and then checked by TLC for substantial completion of the reaction. Ethanol (100. mu.L), saturated sodium carbonate solution (80. mu.L) and 30% hydrogen peroxide solution (100. mu.L) were added to the reaction solution in this order, and the mixture was stirred at room temperature for 2 hours to complete the reaction as detected by TLC. The remaining hydrogen peroxide was neutralized by adding saturated sodium sulfite solution, extracted with ethyl acetate (3 × 30mL), and the combined organic layers were washed with deionized water, 1N dilute hydrochloric acid and saturated brine, respectively, dried over sodium sulfate and concentrated, and subjected to column chromatography to give compound s2023.0 mg (0.037mmol), yield: and (5) 55.3%.¹H NMR(300MHz,CDCl₃) δ 7.36-7.30(m,5H),5.14-5.04(m,2H),4.61(t,1H, J ═ 2.7Hz),3.91-3.87(m,1H),3.80-3.67(m,3H),2.34-2.17(m,4H),2.08(s,3H),2.04-1.03(m, other alicyclic protons), 0.99(s,3H),0.86(s,3H),0.82(s,6H),0.75(s,3H).

(2) Intermediate S20 was subjected to a similar debenzylation procedure to give compound T33(15.5mg, 78.8%).¹H NMR(500MHz,CD₃OD) δ 4.61(t,1H, J ═ 2.5Hz),3.74(dd,1H, J ═ 11.0,4.5Hz),3.66(dd,1H, J ═ 11.0,6.5Hz),3.55(dd,1H, J ═ 11.0,7.5Hz),3.46(dd,1H, J ═ 11.0,7.5Hz),2.58-2.52(m,1H),2.40-2.35(m,1H),2.26-2.22(m,1H),2.06(s,3H),1.98-1.89(m,2H),1.84-1.80(m,1H),1.76-1.74(m,1H),1.68-1.16(m, other fat rings), 1.92 (s, 3.92), 3H (s, 0.85H), 3.85 (s,3H), 3.85H, 3H, 3.85 (m, 3H).

Example 13

(1) Intermediate S17(201mg,0.332mmol) was dissolved in DCM (5mL), DMP (211mg,0.498mmol) was added at 0 deg.C, the temperature was maintained for 15 min, and TLC monitoring indicated that the reaction was complete. Filtered through celite, extracted with ethyl acetate (3X 30mL), and the combined organic layers were washed with deionized water, saturated Na, respectively₂SO₃The aqueous solution and saturated brine are washed, dried and concentrated by sodium sulfate, and separated by column chromatography to obtain a product S2156.0 mg (0.093mmol), the molar yield: 28.0 percent. Intermediate S21 was unstable and was directly dosed to the next step.

(2) Intermediate S21(56.0mg,0.093mmol) and excess 2-methyl-2-butene (0.2mL) were dissolved in t-butanol (2mL) and NaClO was added at room temperature₂(29.5mg,0.326mmol) and NaH₂PO₄(55.8mg,0.465mmol) in water (2mL) was reacted for 4 hours and TLC showed completion. Extracted with ethyl acetate (3X 30mL) and the combined organic layers were separately washed with deionized water, saturated NH₄Washing with a Cl aqueous solution and saturated brine, drying and concentrating the solution through sodium sulfate, and separating the product by column chromatography to obtain S2256.2mg (0.093mmol) and a molar yield: 97.5 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.31(M,5H),5.16-5.05(M,2H),4.62(s,1H),2.80-2.71(M,1H),2.45-2.38(M,1H),2.30-2.12(M,2H),2.07(s,3H),1.92-1.07(M, other aliphatic ring protons), 1.16(d,3H, J ═ 7.2Hz),0.97(s,3H),0.87(s,3H),0.83(s,6H),0.75(s,3H), ESI-MS (M/z):643.4(M + Na)⁺.

(3) Intermediate S22 was subjected to a similar debenzylation procedure to give compound T34(12.4mg, 72.5%).¹H NMR(300MHz,d₆-DMSO) δ 12.02(br s,1H),4.50(s,1H),2.24-2.07(m,3H),2.02(s,3H),1.99-1.07(m, other alicyclic protons), 1.02(d,3H, J ═ 6.9Hz),0.93(s,3H),0.89(s,3H),0.84-0.83(m,6H),0.78(s,3H).

Example 14

(1) Intermediate S17(50.0mg,0.083mmol) was dissolved in dry DMF (3mL) and tris was added at room temperatureThiosyltrimethylamine oxide (150mg,0.830mmol) was heated to 95 ℃ for 24 hours and monitored by TLC to show completion. Diluting with water, extracting with ethyl acetate (3 × 50mL), washing the combined organic layers with deionized water and saturated brine respectively, drying over sodium sulfate, concentrating, and separating by column chromatography to obtain product s2321.5mg (0.031mmol), yield: 37.9 percent.¹H NMR(300MHz,CD₃OD) δ 7.40-7.29(m,5H),5.17-5.05(m,2H),4.59(t,1H, J ═ 2.7Hz),4.16-4.11(m,1H),3.86-3.80(m,1H),2.34-2.14(m,3H),2.05(s,3H),1.99-1.06(m, other alicyclic protons), 1.03(s,3H),0.97(d,3H, J ═ 6.9Hz),0.89(s,3H),0.87(s,3H),0.84(s,3H),0.74(s,3H).

(2) Intermediate S23 was subjected to a similar debenzylation procedure to give compound T35(7.10mg, 70.4%).¹H NMR(300MHz,CD₃OD) δ 4.60(t,1H, J ═ 2.4Hz),4.19-4.14(m,1H),3.87-3.81(m,1H),2.37-2.31(m,2H),2.22(d,1H, J ═ 11.7Hz),2.06(s,3H),2.02-1.14(m, other aliphatic ring protons), 1.06(s,3H),1.00-0.98(m,6H),0.91(s,6H),0.85(s,3H).

Example 15

(1) Intermediate S17(30.0mg,0.050mmol), DMAP (1.22mg,0.010mmol), EDCI (19.1mg,0.100mmol) and 4-morpholinoacetic acid (14.5mg,0.100mmol) were dissolved in dry DCM (2mL) and stirred overnight at room temperature, the next day TLC showed completion of the reaction. The DCM was spun off, extracted with ethyl acetate (3 × 30mL), the combined organic layers were washed with deionized water and saturated brine respectively, dried over sodium sulfate, concentrated, and isolated by column chromatography to give 33.5mg (0.046mmol) of product, yield: 92.0 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.31(m,5H),5.15-5.04(m,2H),4.61(t,1H, J ═ 3.0Hz),4.30-4.25(m,1H),3.90-3.84(m,1H),3.78-3.73(m,4H),3.21(s,3H),2.60-2.57(m,4H),2.40-2.16(m,3H),2.09-2.02(m,4H),1.87-1.80(m,2H),1.67-1.07(m, other fat ring protons), 0.98(s,3H),0.94(d,3H, J ═ 6.9Hz),0.93(s,3H),0.86(s,3H),0.82(s,3H),0.74(s,3H).

(2) Compound T36(22.0mg, 74.9%) was obtained by a similar debenzylation procedure.¹H NMR(500MHz,CDCl₃) δ 4.62(t,1H, J ═ 3.0Hz),4.28(dd,1H, J ═ 10.5,4.5Hz),3.89(dd,1H, J ═ 10.5,8.0Hz),3.76(t,4H, J ═ 4.5Hz),3.23(s,2H),2.62-2.60(m,4H),2.37-2.31(m,1H),2.28-2.18(m,2H),2.09(s,3H),2.07-2.00(m,1H),1.91-1.83(m,2H),1.77-1.09(m, other alicyclic protons), 1.02(s,3H),0.95(d,1H, J ═ 7.0), 0.93(s,3H),0.86(s,3H),0.85(s,3H), 3H, and so on.

In a similar manner to that described in example 15, intermediate S17 was condensed with succinic acid to give compound T37.

Example 16

(1) Intermediate S17(150mg,0.248mmol) was dissolved in dry DCM (2mL), pyridine (99.9. mu.L, 1.24mmol) was added dropwise at 0 deg.C, after 0.5h a solution of p-nitrophenyl chloroformate (150mg,0.744mmol) in dry DCM (3mL) was added dropwise, and after returning to room temperature, the mixture was stirred for 5h, and TLC indicated completion of the reaction. The DCM was spun off, extracted with ethyl acetate (3 × 50mL), the combined organic layers were washed with deionized water and saturated brine respectively, dried over anhydrous sodium sulfate and concentrated, and isolated by column chromatography to give product S24153 mg (0.198mmol) in yield: 79.9 percent.¹H NMR(300MHz,CDCl₃) δ 8.31-8.26(m,2H),7.41-7.31(m,5H +2H),5.16-5.06(m,2H),4.62(t,1H, J ═ 2.7Hz),4.40(dd,1H, J ═ 10.5,4.8Hz),4.07(dd,1H, J ═ 10.5,7.8Hz),2.46-2.19(m,6H),2.06(s,3H),1.91-1.08(m, other fat ring protons), 1.03-1.00(m,6H),0.86(s,3H),0.83(s,3H),0.82(s,3H),0.75(s,3H).

(2) Intermediate S24(40.0mg,0.052mmol), DMAP (19.1mg,0.156mmol) and triethylamine (21.6. mu.L, 0.156mmol) were dissolved in dry DCM (2mL), N- (2-aminoethyl) morpholine (34.0. mu.L, 0.260mmol) was added and stirred at room temperature overnight, the next day TLC showed complete reaction. Diluted with water, extracted with ethyl acetate (3X 30mL), and the combined organic layers were washed with deionized water and saturated brineWashing, drying over sodium sulfate, concentrating, and separating by column chromatography to obtain 35.0mg (0.046mmol) of product, yield: 88.5 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.30(m,5H),5.15-5.05(m,2H +1H),4.61(t,1H, J ═ 2.7Hz),4.18-4.16(m,1H),3.87-3.81(m,1H),3.73-3.70(m,4H),3.32-3.27(m,2H),2.50-2.46(m,6H),2.35-2.13(m,6H),2.08(s,3H),1.92-1.03(m, other fat ring protons), 0.98(s,3H),0.93(d,3H, J ═ 6.6Hz),0.86(s,3H),0.82(s,3H),0.74(s,3H).

(3) Compound T38(24.3mg, 78.7%) was obtained by a similar debenzylation procedure.¹H NMR(300MHz,CDCl₃) δ 5.33(s,1H),4.62(s,1H),4.26-4.21(m,1H),3.82-3.73(m,5H),3.38-3.28(m,2H),3.32-3.27(m,2H),2.52-2.50(m,6H),2.37-2.17(m,6H),2.09(s,3H),2.05-1.10(m, other fat ring protons), 1.02(s,3H),0.98-0.93(m,6H),0.86(s,3H),0.85(s,3H),0.83(s,3H).

Compound T39 was obtained by condensing intermediate S24 with 2-morpholinoethanol in a similar manner to that described in example 16.

Example 17

(1) Intermediate S22(28.0mg,0.045mmol), glycine benzyl ester hydrochloride (10.9mg,0.054mmol), EDCI (13.0mg,0.068mmol), HOBt (6.08mg,0.045mmol) and 4-methylmorpholine (14.8. mu.L, 0.135mmol) were dissolved in dry DMF (2mL) and stirred overnight at room temperature with next day TLC monitoring showing the reaction was complete. After dilution with water and extraction with ethyl acetate (3 × 50mL), the combined organic layers were washed with deionized water and saturated brine, respectively, dried over sodium sulfate, concentrated, and separated by column chromatography to give 27.0mg (0.035mmol) of the product, yield: 77.9 percent.¹H NMR(300MHz,CDCl₃) δ 7.39-7.30(m,10H),5.83(t,1H, J ═ 5.4Hz),5.19(s,2H),5.15-5.04(m,2H),4.62(t,1H, J ═ 3.0Hz),4.08(d,2H, J ═ 5.1Hz),2.51-2.17(m,3H),2.07(s,3H),1.94-1.05(m, other alicyclic protons), 1.1 (s,3H),1.1 (m, other alicyclic protons), and combinations thereof.16(d,3H,J＝6.9Hz),0.95(s,3H),0.87(s,3H),0.83(s,6H),0.74(s,3H).

(2) Compound T40(10.7mg, 51.8%) was obtained by a similar debenzylation procedure.¹H NMR(300MHz,CD₃OD) δ 4.61(s,1H),3.90-3.76(m,2H),2.58-2.54(m,1H),2.35-2.16(m,4H),2.06(s,3H),2.02-1.91(m,1H),1.72-1.12(m, other alicyclic protons), 1.15(d,3H, J ═ 6.9Hz),1.03(s,3H),0.98(s,3H),0.92(s,3H),0.91(s,3H),0.85(s,3H).

Using a similar procedure as in example 17, condensation of intermediate S22 with a different amine gave the following compound.

Example 18

(1) Intermediate S22(81.5mg,0.131mmol), DMAP (1.22mg,0.010mmol), EDCI (37.8mg,0.197mmol) and N- (2-hydroxyethyl) morpholine (159. mu.L, 1.31mmol) were dissolved in dry DCM (2mL) and stirred overnight at RT with the next day of TLC monitoring showing the reaction was complete. DCM was spun off, extracted with ethyl acetate (3 × 50mL), the combined organic layers were washed with deionized water and saturated brine respectively, dried over sodium sulfate and concentrated, and isolated by column chromatography to give 70.2mg (0.096mmol) of product, yield: 72.9 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.29(m,5H),5.15-5.05(m,2H),4.62(t,1H, J ═ 2.7Hz),4.20(t,2H, J ═ 5.7Hz),3.71(t,4H, J ═ 4.5Hz),2.72-2.60(m,4H),2.51(t,4H, J ═ 4.5Hz),2.39-2.12(m,5H),2.08(s,3H),2.04-1.02(m, other alicyclic protons), 1.13(d,3H, J ═ 7.2Hz),0.98(s,3H),0.87(s,3H),0.83(s,6H),0.75(s,3H).

(2) Compound T46(59.3mg, 96.3%) was obtained by a similar debenzylation procedure.¹H NMR(500MHz,CDCl₃) δ 4.62(t,1H, J ═ 3.0Hz),4.23-4.21(m,2H),3.72(t,4H, J ═ 4.5Hz),2.74-2.69(m,1H),2.65(t,2H, J ═ 6.0Hz),2.54(t,4H, J ═ 4.5Hz),2.35-2.31(m,1H),2.27-2.16(m,2H),2.08(s,3H),1.91-1.17(m, other alicyclic protons), 1.14(d,3H, J ═ 7.0Hz),1.01(s,3H),0.94(s,3H),0.87(s,3H),0.86(s,3H),0.84(s,3H).

Example 19

(1) Intermediate S12(1.50g,2.75mmol) was dissolved in dry DCM (20mL), pyridine (665. mu.L, 8.25mmol) was added dropwise at 0 deg.C, after 0.5 hr p-nitrophenyl chloroformate (2.77g,13.8mmol) in dry DCM (10mL) was added dropwise, and after returning to room temperature, stirring was continued for 5 hr, as monitored by TLC showing completion of the reaction. DCM was spun off, extracted with ethyl acetate (3 × 100mL), the combined organic layers were washed with deionized water and saturated brine respectively, dried over anhydrous sodium sulfate and concentrated, and isolated by column chromatography to give product s251.74g (2.45mmol), yield: 89.1 percent.¹H NMR(300MHz,CDCl₃) δ 8.31-8.27(m,2H),7.41-7.33(m,7H),5.17-5.07(m,2H),4.73(d,1H, J ═ 2.4Hz),4.60-4.59(m,1H),3.07-2.98(m,1H),2.31-2.13(m,2H),2.02-1.01(m, other fat ring protons), 0.96(s,3H),0.95(s,3H),0.91(s,3H),0.84(s,3H),0.76(s,3H).

(2) Intermediate S25(100mg,0.141mmol), DMAP (51.7mg,0.423mmol) and triethylamine (58.6. mu.L, 0.423mmol) were dissolved in dry DCM (2mL), cyclopropylamine (58.6. mu.L, 0.910mmol) was added dropwise, stirring overnight at room temperature, the next day TLC showed complete reaction. The DCM was spun off, extracted with ethyl acetate (3X 50mL), and the combined organic layers were washed with deionized water and saturated brine, respectively, dried over sodium sulfate, concentrated, and isolated by column chromatography to give 78mg (0.126mmol) of the product.¹H NMR(300MHz,CDCl₃) δ 7.39-7.33(m,5H),5.18-5.07(m,2H),4.76-4.71(m,2H),4.59(s,1H),4.49(s,1H),3.07-2.98(m,1H),2.81(d,3H, J ═ 3.3Hz),2.32-2.13(m,2H),1.94-1.03(m, other alicyclic protons), 0.97(s,3H),0.85(s,6H),0.81(s,3H),0.76(s,3H).

(3) Compound 22 was prepared by a similar procedure as intermediate S17.4mg(55.3％)。¹H NMR(300MHz,CDCl₃) δ 7.39-7.30(m,5H),5.15-5.05(m,2H),4.64(br s,1H),4.49(s,1H),3.77(dd,1H, J ═ 10.5,4.5Hz),3.42(dd,1H, J ═ 10.5,7.8Hz),2.80(d,3H, J ═ 4.8Hz),2.34-2.16(m,4H),1.92-1.07(m, other fat ring protons), 0.97-0.94(m,6H),0.86-0.82(m,9H),0.74(s,3H).

(4) Compound T47(5.80mg, 96.8%) was obtained by a similar debenzylation procedure.¹H NMR(300MHz,CDCl₃) δ 4.68-4.66(m,1H),4.49(s,1H),3.79(dd,1H, J ═ 10.5,4.8Hz),3.43(dd,1H, J ═ 10.5,8.1Hz),2.80(d,3H, J ═ 4.8Hz),2.35-2.17(m,5H),1.92-1.09(m, other fat ring protons), 0.98-0.96(m,6H),0.93(s,3H),0.86-0.84(m,9H).

In a similar manner to that described in example 19, compounds T48-T50 were obtained.

Example 20

Intermediate S26 product T51(30.7mg, 79.5%) was obtained according to a similar procedure to compound T28.¹H NMR(400MHz,CDCl₃) δ 4.94(br s,1H),4.51(s,1H),3.28-3.22(m,1H),2.58(s,1H),2.28(d,1H, J ═ 10.0Hz),2.15-1.06(m, other alicyclic protons), 1.02(s,3H),0.92(s,3H),0.86-0.84(m,9H),0.74-0.69(m,2H),0.56-0.51(m,2H).

Example 21

Intermediate S26 product T52(42.0mg, 84.2%) was obtained according to a similar procedure to compound T29.¹H NMR(400MHz,CDCl₃) δ 4.92(br s,1H),4.51(s,1H),2.67-2.57(m,3H),2.28(d,1H, J ═ 12.4Hz),2.19-1.02(m, other alicyclic protons), 0.99(s,3H),0.93(s,3H),0.87-0.86(m,9H),0.72-0.71(m,2H),0.54(s,2H).

Example 22

Intermediate S26 product T53(53.0mg, 76.5%) was obtained according to a similar procedure to compound T30.¹H NMR(400MHz,CDCl₃) δ 4.96(br s,1H),4.50(s,1H),3.14(t,1H, J ═ 8.4Hz),2.58(s,1H),2.37(d,1H, J ═ 12.0Hz),2.26 to 1.16(m, other alicyclic protons), 0.96(s,3H),0.91(s,3H),0.84(s,6H),0.80(s,3H),0.71 to 0.70(m,2H),0.54(s,2H).

Example 23

Intermediate S26 product T54(28.2mg, 55.6%) was obtained according to a similar procedure to compound T32.¹H NMR(400MHz,CDCl₃) δ 4.50(s,1H),4.34-4.25(m,2H),3.25-3.18(m,1H),2.57(s,1H),2.32-2.22(m,2H),2.10-1.19(m, other alicyclic protons), 1.00(s,3H),0.90(s,3H),0.85-0.83(m,9H),0.71-0.70(m,2H),0.53(s,2H).

Example 24

Intermediate S26 product T55(7.6mg, 22.4%) was obtained according to a similar procedure to compound T33.¹H NMR(400MHz,CD₃OD) delta 4.41(s,1H),3.70-3.68(m,1H),3.63-3.59(m,1H),3.53-3.48(m,1H),3.44-3.37(m,1H),2.53-2.47(m,2H),2.33-2.32(m,1H),2.21-2.18(m,1H),1.94-1.11(m, other alicyclic protons), 0.99-0.95(m,6H),0.86-0.84(m,9H),0.62(s,2H),0.43(s,2H).

Example 25

Intermediate S26 according to compound T34This was done in analogy to the procedure to give product T56(10.5mg, 57.3%).¹H NMR(300MHz,CD₃OD) δ 4.45(s,1H),2.68-2.65(m,1H),2.51(s,1H),2.38-1.16(m, other alicyclic protons), 1.11(d,3H, J ═ 6.9Hz),0.98(s,6H),0.90-0.87(m,9H),0.67-0.64(m,2H),0.49-0.44(m,2H).

Example 26

(1) Intermediate S27(180mg,0.285mmol) was dissolved in MeOH 5mL and NaBH added at 0 deg.C₄(53.9mg,1.425mmol), the reaction was maintained at this temperature for 5h and TLC showed completion. The reaction was quenched with acetone, the solvent was spun off, extracted with ethyl acetate (3X 50mL), and the combined organic layers were washed with deionized water, saturated Na, respectively₂SO₃The aqueous solution and saturated brine were washed, dried over sodium sulfate, concentrated, and separated by column chromatography to give products S28 and S29, 85.7mg (0.135mmol) and 81.9mg (0.129mmol), respectively, yield: 92.8 percent. S28:¹H NMR(400MHz,CDCl₃) δ 7.36-7.30(m,5H),5.14-5.07(m,2H),4.88(br S,1H),4.50(S,1H),3.89(S,1H),2.58(S,1H),2.30-2.17(m,3H),1.90-1.81(m,3H),1.70-1.08(m, other aliphatic ring protons), 1.13(d,3H, J ═ 6.4Hz),0.96(S,6H),0.82(S,3H),0.75(S,3H),0.72-0.70(m,2H),0.53(S,2H). S29:¹H NMR(400MHz,CDCl₃) δ 7.36-7.31(m,5H),5.11(s,2H),4.84(br s,1H),4.50(s,1H),4.02(s,1H),2.59-2.53(m,2H),2.32-2.29(m,1H),2.22-2.16(m,1H),1.91-1.83(m,2H),1.73-1.10(m, other fat ring protons), 1.06(d,3H, J ═ 6.0Hz),0.92(s,3H),0.86(s,6H),0.82(s,3H),0.74(s,3H),0.72-0.70(m,2H),0.53(s,2H).

(2) Similar debenzylation of intermediates S28 and S29 gave compounds T57(30.2mg, 79.5%) and T58(32.5mg,%). T57:¹H NMR(400MHz,d₆-DMSO) δ 11.91(s,1H),7.15(br s,1H),4.32(s,1H),4.15(d,1H, J ═ 4.8Hz),4.09(q,1H, J ═ 5.2Hz),3.65-3.62(m,1H),2.44(s,1H),2.25-2.18(m,1H),2.10(d,1H, J ═ 12.0Hz),2.05-1.99(m,1H),1.88-1.81(m,1H),1.70-1.64(m,1H),1.56-1.06(m, other alicyclic protons), 0.98(d,3H, J ═ 6.0Hz),0.92(s,3H),0.88(s,3H),0.82(s,9H), 0.9 (s,2H), 2.40H, T (T, 40H), 2H, 40H, T, t.55H, 2H, 1H58:¹H NMR(400MHz,CDCl₃) δ 4.93(br s,1H),4.51(s,1H),4.05(t,1H, J ═ 5.6Hz),2.59-2.54(m,2H),2.29-2.20(m,2H),1.97-1.13(m, other alicyclic protons), 1.08(d,3H, J ═ 6.4Hz),0.94(s,6H),0.87-0.86(m,9H),0.72-0.71(m,2H),0.54(s,2H).

Example 27

(1) Intermediate S30(300mg,0.464mmol) was dissolved in DCM (5mL), DMP (295mg,0.696mmol) was added at 0 deg.C, the temperature was maintained and the reaction was complete as indicated by TLC for 15 min. Filtered through celite, extracted with ethyl acetate (3X 50mL), and the combined organic layers were washed with deionized water, saturated Na, respectively₂SO₃The aqueous solution and saturated brine were washed, dried over sodium sulfate, concentrated, and separated by column chromatography to give product S31210 mg (0.325mmol), yield: 70.1 percent. S31 is unstable and is directly used for the next step.

(2) Intermediate S31(240mg,0.372mmol) was dissolved in dry THF (5mL) and methylmagnesium bromide (186. mu.L, 0.558mmol) was added at-78 deg.C and the temperature was maintained for 15 min and allowed to return to room temperature overnight with TLC indicating completion. Filtered through celite, extracted with ethyl acetate (3X 50mL), and the combined organic layers were washed with deionized water, saturated Na, respectively₂SO₃The aqueous solution and saturated brine were washed, and the concentrated intermediate S32 was dried over sodium sulfate and directly charged to the next step. S32 was subjected to an oxidation procedure similar to S31 to afford intermediate S33 in yields: 81.9 percent.¹H NMR(400MHz,CDCl₃) δ 7.35-7.30(m,5H),5.13-5.05(m,2H),4.90(s,1H),4.50(s,1H),2.87-2.81(m,1H),2.58(s,1H),2.36-2.15(m,3H),2.13(s,3H),2.09(s,1H),2.04(s,1H),1.90-1.06(m, other fatty ring protons), 1.13(d,3H, J ═ 6.8Hz),0.91(s,3H),0.86-0.82(m,9H),0.74(s,3H),0.72-0.70(m,2H),0.53(s,2H).

(4) Intermediate S33 was subjected to a similar debenzylation procedure to give compound T59(40.0mg, 84.2%).¹H NMR(400MHz,CDCl₃) Δ 4.94(s,1H),4.51(s,1H),2.88-2.81(m,1H),2.58(s,1H),2.34-2.19(m,3H),2.14(s,3H),1.90-1.17(m, other alicyclic protons),1.14(d,3H,J＝6.8Hz),0.94(s,3H),0.92(s,3H),0.86-0.85(m,9H),0.72-0.70(m,2H),0.53(s,2H).

Example 28

(1) Esterification of intermediate S34 gave intermediate S35(11.0mg, 31.3%).¹H NMR(300MHz,CDCl₃) δ 7.36-7.29(m,5H),5.15-5.04(m,2H),4.53(t,1H, J ═ 2.7Hz),4.21-4.19(m,2H),3.71(t,4H, J ═ 4.8Hz),2.72-2.55(m,5H),2.51(t,4H, J ═ 4.5Hz),2.41-0.99(m, other fat ring protons), 1.13(d,3H, J ═ 6.9Hz),0.94(s,3H),0.86(s,6H),0.83(s,3H),0.74(s,3H),0.71-0.67(m,2H),0.54-0.49(m,2H).

(2) Intermediate S35 was subjected to a similar debenzylation procedure to give compound T60(7.10mg, 73.0%).¹H NMR(300MHz,CDCl₃) δ 5.22(br s,1H),4.54(s,1H),4.23-4.19(m,2H),3.73(t,4H, J ═ 4.8Hz),2.74-1.04(m, other alicyclic protons), 1.14(d,3H, J ═ 6.9Hz),0.97(s,3H),0.94(s,3H),0.87-0.86(m,9H),0.74-0.68(m,2H),0.54-0.51(m,2H).

Example 29

(1) Reaction of intermediate S36 with morpholineacetic acid gave 28.0mg of condensation product, yield: 75.9 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.30(m,5H),5.14-5.04(m,2H),4.20(dd,1H, J ═ 10.8,5.1Hz),3.91(dd,1H, J ═ 10.8,7.8Hz),3.77-3.74(m,1H),3.38(t,1H, J ═ 2.7Hz),3.21(s,2H),2.61-2.58(m,4H),2.38-1.05(m, other alicyclic protons), 0.94-0.91(m,9H),0.81(s,6H),0.74(s,3H).

(2) Prepared in a similar manner to intermediate S7 to give 95.0mg of the activated carbonate product, yield: 74.5 percent.¹H NMR(300MHz,CDCl₃)δ8.31-8.26(m,2H),7.42-7.31(m,5H+2H),5.15-5.05(m,2H),4.59(s,1H),4.26(dd,1H,J＝10.8,4.5Hz),3.88(dd,1H,J＝10.8,8.1Hz),3.75(t,4H,J＝4.8Hz),3.22(s,2H),2.60-2.57(m,4H),2.36-2.17(m,3H),2.08-105(m, other alicyclic protons), 0.96-0.92(m,12H),0.85(s,3H),0.75(s,3H).

(3) The activated carbonate reacted with cyclopropylamine to give 48.0mg, 98.3% yield.¹H NMR(300MHz,CDCl₃) δ 7.36-7.31(m,5H),5.15-5.05(m,2H),4.51(t,1H, J ═ 2.7Hz),4.27(dd,1H, J ═ 11.1,4.8Hz),3.87(dd,1H, J ═ 10.5,8.4Hz),3.76(t,4H, J ═ 4.8Hz),3.21(s,2H),2.61-2.57(m,5H),2.41-1.06(m, other fat ring protons), 0.95-0.93(m,6H),0.86(s,6H),0.82(s,3H),0.74-0.69(m,3H +2H),0.55-0.53(m,2H).

(4) Compound T66(33.8mg, 79.7%) was obtained by a similar debenzylation procedure.¹H NMR(500MHz,CDCl₃) δ 5.01(br s,1H),4.52(t,1H, J ═ 3.0Hz),4.29-4.23(m,1H),3.89(t,1H, J ═ 9.5Hz),3.76(t,4H, J ═ 4.5Hz),3.22(s,2H),2.61-2.59(m,4H),2.37-2.32(m,1H),2.28-2.19(m,2H),2.05-2.01(m,1H),1.92-1.07(m, other aliphatic ring protons), 0.99(s,3H),0.95(d,3H, J ═ 6.9Hz),0.93(s,3H),0.86-0.85(m,9H),0.73-0.71(m,2H), 0.56-2H (m, 53H).

In a similar manner to example 29, the following compounds were obtained.

Example 30

(1) Intermediate S30(300mg,0.464mmol) was dissolved in dry DCM (5mL), pyridine (187. mu.L, 2.32mmol) was added dropwise at 0 deg.C, after 0.5h p-nitrophenyl chloroformate (280mg,1.39mmol) in dry DCM (5mL) was added dropwise, and after returning to room temperature, stirring was continued for 5h, as monitored by TLC showing completion of the reaction. The DCM was spun off, extracted with ethyl acetate (3 × 50mL), the combined organic layers were washed with deionized water and saturated brine respectively, dried over anhydrous sodium sulfate and concentrated, and isolated by column chromatography to give product S37374 mg (0.461mmol) in yield: 99.5 percent.¹H NMR(300MHz,CDCl₃)δ8.32-8.26(m,2H),7.40-7.30(m,7H),5.16-5.06(m,2H),4.85(br s,1H),4.50(s,1H),4.42-4.37(m,1H),4.11-4.04(m,1H),2.56(s,1H),2.46-2.39(m,1H),2.32-2.16(m,3H),1.91-1.07(m, other aliphatic ring protons), 1.02(d,3H, J ═ 6.9Hz),0.96(s,3H),0.86-0.83(m,9H),0.75(s,3H),0.71-0.65(m,2H),0.53-0.48(m,2H).

(2) Condensation of intermediate S37 with N- (3-aminopropyl) morpholine gave 14.5mg, 48.0% yield.¹H NMR(400MHz,CDCl₃) δ 7.37-7.31(m,5H),5.56(br s,1H),5.23(br s,1H),5.14-5.05(m,2H),4.51(t,1H, J ═ 2.8Hz),4.19-4.17(m,1H),3.81-3.71(m,5H),3.30-3.25(m,2H),2.59(s,1H),2.45-2.42(m,6H),2.37-1.06(m, other aliphatic ring protons), 0.95-0.92(m,6H),0.86-0.79(m,9H),0.73-0.70(m,3H +2H),0.54-0.50(m,2H).

(3) Compound T75(10.1mg, 78.3%) was obtained by a similar debenzylation procedure.¹H NMR(400MHz,CDCl₃) δ 5.49(br s,1H),5.21(s,1H),4.51(s,1H),4.19(s,1H),3.82-3.73(m,5H),3.27-3.24(m,2H),2.59(s,1H),2.49-2.45(m,6H),2.35-2.30(m,1H),2.26-2.20(m,2H),1.99(s,1H),1.89-1.84(m,2H),1.72-1.09(m, other fat ring protons), 0.98(s,3H),0.94-0.92(m,6H),0.86-0.84(m,9H),0.72-0.69(m,2H),0.56-0.51(m,2H).

In a similar manner to example 30, the following compounds were obtained.

Example 31

(1) Intermediate S7(100mg,0.140mmol) was dissolved in 2mL of DCM with 2, 2' - (ethylenedioxy) bis (ethylamine) (10.4mg,0.070mmol), triethylamine (19.4. mu.L, 0.140mmol) and DMAP (1.71mg,0.014mmol), stirred overnight at room temperature and TLC monitoring indicated that the reaction was complete. The DCM was spun off, diluted with 30mL of deionized water, extracted with ethyl acetate (3 × 30mL), the combined organic layers were washed with deionized water and saturated brine respectively, dried over anhydrous sodium sulfate, concentrated, and isolated by column chromatography to give the product, s3851.6 mg (0.040mmol), yield: 28.6 percent.¹H NMR(300MHz,CDCl₃) δ 7.38-7.30(m,10H),5.21(br s,2H),5.15-5.06(m,4H),4.49(s,2H),3.65(s,4H),3.61-3.57(m,4H),3.40(s,4H),2.28-1.03(m, other alicyclic protons), 0.95(s,6H),0.85-0.81(m,24H),0.75-0.72(m,12H).

(2) Compound T89(15.1mg, 34.0%) was obtained by a similar debenzylation procedure.¹H NMR(300MHz,CDCl₃) δ 5.18(br s,2H),4.51(s,2H),3.66(s,4H),3.62-3.58(m,4H),3.40(s,4H),2.28-1.07(m, other alicyclic protons), 1.00(s,6H),0.93(s,6H),0.86-0.85(m,24H),0.77-0.74(m,6H).

Example 32

(1) Intermediate S37(100mg,0.123mmol) was dissolved in 2mL of DCM with 2, 2' - (ethylenedioxy) bis (ethylamine) (9.2mg,0.062mmol), triethylamine (17.1. mu.L, 0.123mmol) and DMAP (1.47mg,0.012mmol) and stirred overnight at room temperature with TLC monitoring showing the reaction was complete. After the DCM was separated by centrifugation, diluted with 30mL of deionized water, extracted with ethyl acetate (3 × 30mL), and the combined organic layers were washed with deionized water and saturated brine, respectively, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to give s 3966.7 mg (0.045mmol), yield: 36.6 percent.¹H NMR(300MHz,CDCl₃) δ 7.36-7.32(m,10H),5.15-5.05(m,4H),4.51(s,2H),3.63(s,4H),3.57(t,4H, J ═ 5.1Hz),3.39(s,4H),2.39-1.04(m, other alicyclic protons), 0.93(s,12H),0.86 (m,12H), and so on(s,12H),0.82(s,6H),0.73-0.70(m,10H),0.55-0.54(m,4H).

(2) Compound T90(32.4mg, 55.2%) was obtained by a similar debenzylation procedure.¹H NMR(300MHz,CDCl₃) δ 4.50(s,2H),3.63(s,4H),3.57(t,4H, J ═ 5.1Hz),3.40-3.38(m,4H),2.60(s,2H),2.38-1.13(m, other alicyclic protons), 0.97-0.92(m,18H),0.86-0.84(m,18H),0.74-0.68(m,4H),0.56-0.51(m,4H).

Example 33

(1) Reaction of intermediate S29 with morpholineacetic acid gave 19.2mg of condensation product, yield: 45.6 percent.¹H NMR(400MHz,CDCl₃) δ 7.34-7.30(m,5H),5.18-5.06(m,3H),4.85(br s,1H),4.50(s,1H),3.73-3.61(m,3H),3.14(s,1H),2.65-2.57(m,5H),2.31-1.05(m, other alicyclic protons), 0.91(s,3H),0.86(s,6H),0.82(s,3H),0.72-0.69(m,5H),0.53(s,2H).

(2) Compound T91(13.2mg, 77.9%) was obtained by a similar debenzylation procedure.¹H NMR(400MHz,CDCl₃) δ 5.14(t,1H, J ═ 4.8Hz),4.41(s,1H),3.66(s,4H),3.16(s,2H),2.63-2.47(m,6H),2.31-1.18(m, other alicyclic protons), 1.13(d,3H, J ═ 6.4Hz),0.94(s,6H),0.88-0.84(m,9H),0.62(s,2H),0.42(s,2H).

Example 34

(1) Intermediate S29(100mg,0.158mmol) and DMAP (193mg,1.58mmol) were dissolved in dry DCM (2mL) and after 0.5h a solution of p-nitrophenyl chloroformate (63.7mg,0.316mmol) in dry DCM (2mL) was added dropwise, the mixture was allowed to return to room temperature and stirred for 5h, after which time TLC monitoring indicated completion of the reaction, followed by addition of N- (3-aminopropyl) morpholine (231. mu.L, 1.58mmol) dropwise and reaction overnight at room temperature. DCM was spun off, extracted with ethyl acetate (3X 50mL), the combined organic layers were washed with deionized water and saturated brine, respectively, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to give 91.2m productg (0.128mmol), yield: 81.0 percent.¹H NMR(400MHz,CDCl₃) δ 7.35-7.30(m,5H),5.58(s,1H),5.14-5.06(m,2H),4.99(br s,1H),4.86(br s,1H),4.51(s,1H),3.71(s,4H),3.25-3.23(m,2H),2.64-2.58(m,2H),2.45(s,6H),2.31-2.28(m,1H),2.20-2.14(m,1H),1.89-0.96(m, other fat ring protons), 0.91(s,3H),0.86(s,6H),0.81(s,3H),0.72-0.70(m,5H),0.53(s,2H).

(2) Compound T94(50.2mg, 55.0%) was obtained by a similar debenzylation procedure.¹H NMR(400MHz,CD₃OD) δ 4.96(s,1H),4.45(s,1H),3.71(s,4H),3.13(s,2H),2.66-2.61(m,1H),2.55-2.45(m,7H),2.37-2.31(m,1H),2.25(d,1H, J ═ 12.4Hz),1.97-1.12(m, other alicyclic protons), 0.97(s,6H),0.91-0.88(m,9H),0.65(s,2H),0.47(s,2H).

In a similar manner to example 34, the following compounds were obtained.

Example 35

(1) Intermediate S30(50mg,0.0731mmol) and triphenylphosphine (48mg, 0.183mmol) were dissolved in dry THF (2mL), diisopropyl azodicarboxylate (75. mu.L, 0.381mmol) was added dropwise at 0 deg.C, diphenyl azidophosphate (75mg,0.272mmol) was added dropwise after 15 minutes, the mixture was allowed to return to room temperature and stirred overnight, TLC monitored to show completion, a solution of triphenylphosphine (200mg, 0.762mmol) in THF (2mL) was added and stirred overnight, TLC monitored to show completion, water (2mL) was added, stirring was allowed 4 hours, and TLC monitored to show completion. THF was then decanted, extracted with ethyl acetate (3X 50mL), and the combined organic layers were washed with deionized water and saturated brine, respectively, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatographyProduct S4029.2mg (0.0452mmol) was isolated, yield: 61.8 percent.¹H NMR(400MHz,CDCl₃) δ 7.35-7.30(m,5H),5.09(q,2H, J ═ 12.0Hz),4.92(s,1H),4.50(s,1H),2.91(d,4H, J ═ 12.0Hz),2.58(s,1H),2.42(t,1H, J ═ 11.2Hz),2.34(t,1H, J ═ 10.8Hz),2.30-2.26(m,1H),2.22(d,1H, J ═ 12.0Hz),2.17(s,1H), 1.86-1.20 (m, other fat ring protons), 1.08(d,3H, J ═ 10.8Hz),0.98(d,3H, J ═ 6.8Hz),0.94(s,3H), 0.86(s,8H), 0.81(s,4H, J ═ 6.53, H, 0.53 (d, H), 2H, J ═ 6.0.53 Hz).

(2) Compound T96(16.7mg, 66.4%) was obtained by a similar debenzylation procedure.¹H NMR(400MHz,CD₃OD) δ 4.45(s,1H),3.11(d,1H, J ═ 11.2Hz),2.69(t,1H, J ═ 12.0Hz),2.52-2.46(m,3H),2.27(d,1H, J ═ 12.8Hz),2.05-1.16(m, other alicyclic protons), 1.06(d,3H, J ═ 6.8Hz),1.03-0.99(m,8H),0.98(s,2H), 0.92(s,7H),0.89(s,3H),0.67(s,2H),0.48(s,2H).

Example 36

(1) Intermediate S40(50mg,0.0773mmol) was dissolved in dry DCM (2mL), 4-dimethylaminopyridine (9.44mg,0.0773mmol) and triethylamine (31.49. mu.L, 0.386mmol) were added, and after 5 minutes a solution of p-nitrophenyl chloroformate (46.73mg,0.232mmol) in dry DCM (1mL) was added dropwise, stirred at RT for 5 hours, and after TLC monitoring indicated that the reaction was complete, 3- (4-morpholine) -1-propanol (50.00. mu.L, 0.386mmol) was added and stirred at RT overnight. TLC monitoring indicated complete reaction. DCM was spun off, extracted with ethyl acetate (3 × 50mL), the combined organic layers were washed with deionized water and saturated brine respectively, dried over anhydrous sodium sulfate and concentrated, and isolated by column chromatography to give the product s4138.1mg (0.0524mmol) in yield: 60.3 percent.¹H NMR(400MHz,CDCl₃) δ 7.35-7.31(m,5H),5.09(q,2H, J ═ 12.0Hz),4.51(s,1H),4.21(s,1H),4.12-4.08(m,1H),3.73(t,3H, J ═ 4.8Hz),3.39(s,1H),2.76(s,1H),2.60(d,1H, J ═ 10.4Hz),2.47-2.42(m,4H),2.35(d,1H, J ═ 11.2Hz),2.29-2.26(m,1H), 2.23-2.16 (m,1H),1.85-1.25(m, other fat ring protons), 1.08(d,3H, J ═ 12.4), 0.96(s,2H),0.91(d,3H, J ═ 12.4), 0.83(s, 2H), 3H, 7(d,3H, 7H, 3H, 12H, d ═ 12H (d,3H,J＝15.6Hz),0.73(s,3H),0.71(d,2H,J＝6.8Hz),0.53(s,2H).

(2) compound T97(27.6mg, 72.4%) was obtained by a similar debenzylation procedure.¹H NMR(400MHz,CDCl₃) δ 4.52(s,1H),4.09(d,1H, J ═ 6.4Hz),3.75(t,3H, J ═ 4.8Hz),3.40(s,1H),2.78(s,1H),2.60(s,1H),2.50-2.46(m,5H),2.32-2.20(m,4H),2.07-2.00(m,2H),1.86-1.08(m, other fat ring protons), 0.99(d,6H, J ═ 6.8Hz),0.92(d,7H, J ═ 6.0Hz),0.87-0.84(m,13H),0.72(d,2H, J ═ 6.8Hz),0.54(s,2H).

In a similar manner to that described in example 36, Compound T98 was obtained.

Example 37

(1) Oleanolic acid (200mg,0.439mmol) was dissolved in DMF (5mL), anhydrous potassium carbonate (121mg,0.878mmol) was added, and benzyl chloride (60.7. mu.L, 0.527mmol) was slowly added dropwise with stirring at room temperature. After the completion of the dropwise addition, the reaction solution was transferred to 50 ℃ and stirred for 3 hours, and the completion of the reaction was monitored by TLC. The mixture was cooled to room temperature, diluted with 50mL of deionized water, extracted with ethyl acetate (3 × 50mL), and the combined organic layers were washed with deionized water and saturated brine, respectively, dried over anhydrous sodium sulfate, concentrated, and isolated by column chromatography to give product S43208 mg (0.381mmol), yield: 86.9 percent.¹H NMR(300MHz,CDCl₃) δ 7.38-7.28(m,5H),5.29(t,1H, J ═ 3.9Hz),5.12-5.02(m,2H),3.23-3.18(m,1H),2.94-2.88(m,1H),2.04-1.93(m,1H),1.89-1.83(m,1H),1.72-1.22(m, other fat ring protons), 1.13(s,3H),0.98(s,3H),0.92(s,3H),0.90(s,3H),0.88(s,3H),0.78(s,3H),0.61(s,3H).

(2) S43(208mg,0.381mmol) was dissolved in DCM (5mL), Dess-Martin oxidant (323mg,0.762mmol) was added slowly at 0 deg.C, allowed to warm slowly to room temperature and stirred for 1 hour, and the reaction was shown to be complete by TLC monitoring. The reaction mixture was filtered and spin-dried overColumn chromatography gave compound S44190 mg (0.349mmol), yield: 91.7 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.28(m,5H),5.31(t,1H, J ═ 3.6Hz),5.13-5.03(m,2H),2.92(dd,1H, J ═ 13.8,4.5Hz),2.60-2.48(m,1H),2.40-2.31(m,1H),2.04-1.17(m, other alicyclic protons), 1.13(s,3H),1.08(s,3H),1.04(s,3H),1.02(s,3H),0.92(s,3H),0.90(s,3H),0.66(s,3H).

(3) The product of the above step, S44(190mg,0.349mmol) and S- (-) -2-methyloxazaborolidine (19.4mg,0.070mmol) were dissolved in dry THF (10 mL). Slowly dropping 10M BH at room temperature₃-Me₂S (41.9. mu.L, 0.419mmol), stirred for 10 min and TLC monitored to show completion of the reaction. Moving the reaction bottle to an ice water bath, slowly dropwise adding methanol to quench the reaction, drying the solvent after no bubbles are generated, and separating by column chromatography to obtain a compound S4540.1mg (0.073mmol) with the yield: 21.0 percent.¹H NMR(300MHz,CDCl₃) δ 7.37-7.32(m,5H),5.29(t,1H, J ═ 3.6Hz),5.12-5.02(m,2H),3.40(t,1H, J ═ 3.0Hz),2.94-2.88(m,1H),1.99-1.84(m,3H),1.72-1.17(m, other alicyclic protons), 1.14(s,3H),0.95(s,3H),0.92(s,3H),0.90-0.89(m,6H),0.84(s,3H),0.62(s,3H).

(4) S45(153mg,0.280mmol) and DMAP (2.44mg,0.02mmol) were dissolved in pyridine (5mL), acetic anhydride (132. mu.L, 1.40mmol) was added dropwise at 0 ℃ and after 10 min returned to room temperature stirring was overnight, the next day TLC showed complete reaction. The pyridine was spun off, extracted with ethyl acetate (3 × 50mL), and the combined organic layers were washed with deionized water, 1N dilute hydrochloric acid and saturated brine, respectively, dried over sodium sulfate, concentrated, and separated by column chromatography to give product S46163 mg (0.277mmol), yield: 98.9 percent.¹H NMR(300MHz,CDCl₃) δ 7.35-7.30(m,5H),5.30(t,1H, J ═ 3.6Hz),5.12-5.02(m,2H),4.63(t,1H, J ═ 3.0Hz),2.91(dd,1H, J ═ 13.8,4.5Hz),2.06(s,3H),2.00-0.96(m, other alicyclic protons), 1.18(s,3H),0.92(s,3H),0.90(s,6H),0.88(s,3H),0.84(s,3H),0.62(s,3H).

(5) Intermediate S46 was subjected to a similar debenzylation procedure to give compound T99(129mg, 93.4%).¹H NMR(300MHz,CDCl₃)5.30(t,1H, J ═ 3.6Hz),4.63(t,1H, J ═ 3.0Hz),2.83(dd,1H, J ═ 14.1,4.5Hz),2.07(s,3H),2.04-1.07(m, other alicyclic protons), 1.19(s,3H),0.93(s,6H),0.91(s,3H),0.88(s,3H),0.85(s,3H),0.77(s,3H).

in a similar manner to example 37, the following compounds were obtained.

Example 38

(1) Chromium trioxide (13.6mg,0.136mmol) was dissolved in dry DCM (1.0mL), pyridine (21.9. mu.L, 0.272mmol) was added dropwise at 0 deg.C and stirred for 1 hour, then a solution of intermediate S47(40mg,0.068mmol) in dry DCM (1.0mL) was added and stirred overnight at room temperature, the next day about half of the starting material remained. Filtering, directly concentrating, separating by column chromatography to obtain product S488.7mg, recovering raw material 29mg, yield: 77.3 percent.¹H NMR(300MHz,CDCl₃) δ 7.35-7.29(m,5H),5.59(s,1H),5.04(s,2H),4.63(t,1H, J ═ 3.0Hz),2.58-2.51(m,1H),2.44(d,1H, J ═ 11.1Hz),2.37(s,1H),2.06(s,3H),2.15-1.06(m, other fat ring protons), 1.33(s,3H),1.10(s,3H),0.97-0.84(m,12H),0.76(s,3H).

(2) Intermediate S48 was subjected to a similar debenzylation procedure to give compound T101(5.6mg, 75.7%).¹H NMR(300MHz,CDCl₃) δ 5.59(s,1H),4.63(t,1H, J ═ 3.0Hz),2.58-2.53(m,1H),2.40(s,1H),2.36(s,1H),2.06(s,3H),2.13-1.09(m, other alicyclic protons), 1.35(s,3H),1.14(s,3H),0.98-0.91(m,6H),0.88-0.86(m,6H),0.84(s,3H).

Example 39

(1) Intermediate S47(59.8mg,0.102mmol) was dissolved in DCM (1.0mL), excess formic acid (100. mu.L) was added at room temperature, followed by the addition of hydrogen peroxide (30%, 52. mu.L, 0.408mmol) and stirring overnight at room temperature, the next day with half of the starting material remaining on the plate. Adding anhydrous sodium sulfiteQuenching, whirling out DCM, extracting with ethyl acetate, washing with water, concentrating the organic phase, separating by column chromatography to obtain product S4921.1mg (0.035mmol), recovering 37.2mg of raw material, yield: 91.4 percent.¹H NMR(300MHz,CDCl₃) δ 7.38-7.31(m,5H),5.22-5.09(m,2H),4.64(t,1H, J ═ 2.7Hz),2.85-2.78(m,1H),2.64-2.54(m,1H),2.43-2.36(m,1H),2.06(s,3H),1.93-1.00(m, other alicyclic protons), 1.33(s,3H),0.96-0.91(m,9H),0.85(s,3H),0.70-0.66(m,6H).

(2) Intermediate S49 was subjected to a similar debenzylation procedure to give compound T102(17.1mg, 95.2%).¹H NMR(300MHz,CDCl₃) δ 4.65(t,1H, J ═ 2.7Hz),2.76-2.70(m,1H),2.64-2.54(m,1H),2.43-2.37(m,1H),2.07(s,3H),1.93-1.08(m, other alicyclic protons), 1.31(s,3H),1.02(s,3H),0.95(s,3H),0.91(s,3H),0.86(s,3H),0.81(s,3H),0.73(d,3H, J ═ 5.4Hz).

Example 40

(1) Intermediate S45(89.0mg,0.163mmol) was dissolved in dry DCM (1mL), pyridine (65.7. mu.L, 0.815mmol) was added dropwise at 0 deg.C, after 0.5h p-nitrophenyl chloroformate (98.6mg,0.489mmol) in dry DCM (1mL) was added dropwise, and after returning to room temperature, the mixture was stirred for 5h, and TLC indicated completion of the reaction. The DCM was spun off, extracted with ethyl acetate (3 × 50mL), the combined organic layers were washed with deionized water and saturated brine respectively, dried over anhydrous sodium sulfate and concentrated, and isolated by column chromatography to give the product S50102 mg (0.143mmol), yield: 88.0 percent.¹H NMR(300MHz,CDCl₃) δ 8.29-8.23(m,2H),7.39-7.31(m,5H +2H),5.29(t,1H, J ═ 3.6Hz),5.12-5.02(m,2H),4.60(t,1H, J ═ 3.0Hz),2.91(dd,1H, J ═ 13.8,4.5Hz),2.03-1.02(m, other alicyclic protons), 1.14(s,3H),0.97(s,3H),0.93-0.91(m,12H),0.61(s,3H).

(2) Condensation of intermediate S50 with cyclopropylamine gave 14.0mg, yield: 69.8 percent.¹H NMR(300MHz,CDCl₃) δ 7.36-7.29(m,5H),5.30(t,1H, J ═ 3.6Hz),5.12-5.02(m,2H),4.82(br s,1H),4.52(t,1H, J ═ 2.7Hz),2.91(dd,1H, J ═ 14.1,4.5Hz),2.57(s,1H),2.04-1.02(m, other fats)Ring protons), 1.15(s,3H),0.92-0.88(m,15H),0.73-0.69(m,2H),0.62(s,3H),0.55-0.50(m,2H).

(3) Compound T105(7.20mg, 60.0%) was obtained by a similar debenzylation procedure.¹H NMR(300MHz,CDCl₃)5.29(t,1H, J ═ 3.3Hz),4.95(br s,1H),4.52(t,1H, J ═ 2.7Hz),2.83(dd,1H, J ═ 13.8,4.5Hz),2.57(s,1H),2.03-1.05(m, other alicyclic protons), 1.15(s,3H),0.93-0.88(m,15H),0.76(s,3H),0.71-0.69(m,2H),0.55-0.50(m,2H).

In a similar manner to example 40, the following compounds were obtained.

EXAMPLE 41

Compound T109 was obtained starting from intermediate S51 via a synthetic procedure similar to compound T31.¹H NMR(400MHz,d₆-DMSO) δ 11.98(br s,1H),4.38(dd, J ═ 11.6,4.8Hz,1H),4.24(t,1H, J ═ 5.2Hz),3.58-3.53(m,1H),3.17-3.10(m,1H),2.25-2.08(m,3H),2.00(s,3H),1.70-1.21(m, other alicyclic protons), 1.07(d,3H, J ═ 11.2Hz),0.93(s,3H),0.87-0.79(m,15H).

Example 42

Compound T110 was obtained starting from intermediate S51 via a synthetic procedure similar to compound T49.¹H NMR(400MHz,d₆-DMSO) delta 11.97(br s,1H),7.14(br s,1H),4.25-4.18(m,2H),3.58-3.53(m,1H),3.17-3.10(m,1H),2.44(s,1H),2.25-2.08(m,3H),1.70-1.23(m, other fats)Cyclic protons), 1.07(d,3H, J ═ 11.6Hz),0.93(s,3H),0.87-0.76(m,15H),0.55-0.52(m,2H),0.37-0.36(m,2H).

Example 43TGR5 receptor agonism assay

1. Purpose of experiment

HEK293 cells using transient TGR5 were stimulated with compounds, and Homogeneous Time-Resolved Fluorescence (HTRF) was used to determine whether these compounds could agonize TGR 5.

2. Principle of experiment

TGR5 is a bile acid membrane receptor, also a member of the GPCR family, and has a regulatory role in the metabolism of bile acids, lipids and carbohydrates. TGR5 is coupled to Gs protein and, after activation, further activates adenylate cyclase, producing the second messenger cAMP. HTRF is a method for detecting cAMP content, which combines two technologies of Fluorescence Resonance Energy Transfer (FRET) and Time Resolved Fluorescence (TRF). The Eu-containing cryptate is used as a fluorescence donor, the emission spectrum of the Eu-containing cryptate overlaps with the excitation spectrum of a fluorescence acceptor to a certain extent, fluorescence is generated by the fluorescence acceptor through FRET induction, the fluorescence life of Eu is long, and a fluorescence signal emitted by the acceptor can be distinguished from a fluorescence background through TRF. The fluorescence donor is bound to the cAMP-specific antibody while cAMP is labeled with a fluorescence acceptor, the two are brought close to each other by an antigen-antibody specific recognition reaction to generate FRET, and cAMP produced by cells competes with the labeled cAMP for an antibody binding site, resulting in a decrease in fluorescence intensity. This experiment uses TGR5 agonist INT777 as a positive control to explore the effect of compounds on TGR 5.

3. Experimental sample

Before the test, the compound is dissolved in DMSO to prepare a mother solution, and the mother solution is diluted to a required concentration by using a culture solution.

4. Experimental methods

4.1 test compounds were made 2-fold higher with 1xPBS at final concentrations of 100. mu.M, 10. mu.M, 1. mu.M, 100nM, 10nM, 1nM, 0.1nM, DMSO (1% DMSO in each well).

4.2 cell treatment:

(1) cells were digested with pancreatin and then suspended in serum-free medium.

(2) The cell density was determined and IBMX (final concentration 500. mu.M) was added to the serum-free medium at the same time, and the cell count was 2000/5. mu.l/well.

(3) Adding 5 mu L of the compound to be detected and 5 mu L of the cell suspension containing IBMX, mixing, sealing the 384-hole plate by using tinfoil paper, and reacting for no more than 30 minutes at room temperature in a dark place.

4.3 assay substrate configuration

(1) mu.L of cAMP-d2 was diluted to 20. mu.L with cAMP & cGMP conjugates & lysine buffer.

(2) mu.L of anti-cAMP-Cryptotate was diluted to 20. mu.L with cAMP & cGMP conjugates & lysine buffer.

(3) After 30 minutes, 5. mu.L (1.3.1) + 5. mu.L (1.3.2) were added, the 384-well plate was sealed with tinfoil, and the reaction was carried out for 30 minutes at room temperature in the dark.

After 4.460 minutes, Envision2101 multifunctional microplate reader (PerkinElmer) readings.

5. Results of the experiments (40 compounds such as T2 are exemplified but not limited to)

Table 1 compound TGR5 agonist activity test assay

Note: EC (EC)₅₀For evaluation of the sample drugs for TGR5 agonist activity, half 50% effective concentration. 1-10 μ M: ". x", 0.1-1 μ M: "x", 100-1 nM: "***"

6. Results and discussion:

as can be seen from the TGR5 receptor agonistic activity test results in table 1, most of the compounds disclosed in the present invention have significantly higher agonistic activity of the humanized TGR5 receptor than that of INT777 and the compounds disclosed in WO2015135449, and as can be seen from the humanized TGR5 agonistic activity results of T109 and T110, the α -configuration compound is significantly better than the β -configuration compound.

EXAMPLE 44 Compound kinetic solubility test

1. Principle of experiment

The precipitation of the compound is used as an indicator of the limitation of the concentration of the compound. The presence of precipitation of the compound is detected indirectly by the increase in the absorption of ultraviolet light by the solution, since the particles block the light from reaching the detector, so that the intensity of the light is reduced.

2. Experimental procedure

The compound solid powder was made into DMSO solution. A small amount of compound DMSO solution was added to DPBS buffer at pH 2.0 and pH7.4, respectively, until the upper concentration limit was reached. The precipitate that appears when the upper concentration limit is reached, the solubility at which the precipitate appears, i.e. the kinetic solubility value, is detected by optical methods.

3. Results of the experiment

TABLE 2 kinetic solubility of the Compounds

4. Results and discussion

The kinetic solubility results are shown in table 2: the kinetic solubility of the compounds T36, T38, T39, T64 and T66 is improved compared with that of C101.

EXAMPLE 45 Compound Caco-2 monolayer cell Permeability assay

1. Experimental procedures

1.1 Caco-2 cell culture

The Caco-2 cells were cultured in high-glucose DMEM at 37 ℃ and 5% CO₂Culturing in an incubator with air relative humidity of 90%, and adding 10% fetal calf serum, 10mmol/L HEPES, 1mmol/L sodium pyruvate, 1% glutamine, 1% nonessential amino acids, 100U/mL penicillin and 100 μ g/mL streptomycin into the culture medium. Passage rate was 1:10 every 7 days. The experiments used between 40 and 60 passages of cells.

1.2 Caco-2 cell monolayer model establishment

Caco-2 cells were plated at 2X 10⁵The cells were inoculated in a Millicell-24 well plate at a concentration of 400. mu.L/well, and 800. mu.L of the culture broth was added to the basal side,at 37 ℃ 5% CO₂Cultured in an incubator. After 72 hours of cell inoculation, the solution was changed, every other day, and the cells were cultured for 21 days.

1.3 Caco-2 cell monolayer model validation

After 21 days of culture, the degree of tightness of the cell monolayer, generally greater than 200. omega. cm, was measured by TEER values²The single layer can be regarded as compact and complete. Higher TEER values indicate a denser monolayer, generally not exceeding 1000. omega. cm². Caco-2 cells were cultured in Millicell culture plates and transmembrane resistance was measured at 21 days using a resistance meter, Millicell-ERS II.

1.4 bilateral transport assay for test Compounds

Drug transport from the top layer (side a) of the cell to the basal layer (side B) and side B to side a was examined. The test method is as follows: after washing the cells three times with HBSS, the corresponding concentrations of compounds were added to the corresponding wells (pH6.8 on the A side and pH7.4 on the B side). Incubate at 37 ℃ for 95 minutes in an incubator, with dosing side sampling at 5 minutes and 95 minutes, and receiving side sampling at 35 minutes and 95 minutes, respectively. The concentration of the sample was measured by LC-MS/MS. To validate cells, a standard curve was fitted using the expected Fabs and Papp values for several positive control compounds. The Papp value of the test compound is determined and the Fabs value is calculated from the standard curve.

2. Results of the experiment

TABLE 3 results of Caco-2 monolayer cell permeability experiments

a：P_app<2×10^-6cm/s low permeability; 2X 10^-6<P_app<20×10^-6cm/s medium permeability; p_app>20×10^-6cm/s high permeability

3. Results and discussion

The results are shown in Table 3: compound C101(WO2015135449,

the existence of the compound can not be detected at all on the apical membrane side or basal membrane side of the Caco-2 monolayer cells, which indicates that the compound can not permeate the cell membrane, the compound can be directly adhered to the cell membrane due to high fat solubility, or the compound can enter the cells and can not be exuded; the permeability of the compounds T36, T38, T39, T64, T66 and T75 which introduce large polar groups into the molecule is obviously improved. The permeation rates of the compounds T36, T38, T39 and T64 are similar on two sides of the membrane, and the permeation rates of T66 and T75 are obviously different on two sides of the membrane, which indicates that T66 and T75 are probably substrates of Pgp protein.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims

1. A compound of formula I, a pharmaceutically acceptable salt thereof, or a dimer thereof:

R₁is hydrogen, hydroxy, halogen or C1-C6 alkyl;

R₂is hydrogen, hydroxy, carboxy, halogen, C1-C6 alkoxy, 3-to 10-membered ring alkoxy, ═ O, ═ N-OH or R-Y (═ X)_m-O-; wherein m is 1 or 2; r is the following substituted or unsubstituted group: C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, 5-7 membered saturated heterocycle, R_aN (C1-C6 alkyl) -, R_aNH-or R_aO-; wherein each R is_aIndependently selected from the group consisting of substituted or unsubstituted: hydrogen, C1-C8 alkanesA C3-C10 cycloalkyl group, a C6-C10 aryl group, a 5-7 membered heteroaryl group, or a 5-7 membered saturated heterocyclic group; x is NH, O or S, Y is C, S or P;

R₅hydrogen, hydroxy, hydroxymethyl, formyl or carboxy;

R₆and R₇Each independently hydrogen, hydroxy, or C1-C6 alkyl;

R₁₀is hydrogen, hydroxy, oxo (═ O), or C1-C6 alkyl;

R₁₁is hydrogen, hydroxy, oxo (═ O), or C1-C6 alkyl;

z is- (CH)₂) n-, n is 1, 2 or 3;

represents a single bond or a double bond;

each independently represents the R configuration, S configuration or racemic;

the substitution means that the hydrogen on the group is substituted by one or more substituents selected from the group consisting of: hydroxyl, amino, methoxy, halogen, C1-C6 alkyl, C1-C6 alkoxy, oxo, epoxy, oxime, carboxyl, sulfonic, glucosamine, morpholinyl, N-dimethyl, 4-methylpiperazinyl, pyridyl, piperidinyl, pyrrolidinyl.

2. The compound of claim 1, wherein R is₁Is hydrogen, hydroxy or C1-C4 alkyl.

3. The compound of claim 1, wherein R is₂Is hydrogen, hydroxy, carboxy, halogen, C1-C4 alkoxy, 3-to 6-membered ring alkoxy, ═ O, ═ N-OH or R-Y (═ X)_m-O-；

Wherein m is 1 or 2;

r is the following substituted or unsubstituted group: C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, 5-7 membered saturated heterocycle, R_aN (C1-C4 alkyl) -, R_aNH-or R_aO-; wherein each R is_aIndependently substituted or unsubstituted: hydrogen, C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, or 5-7 membered saturated heterocyclyl;

x is NH, O or S;

y is C, S or P;

the substitution means that the hydrogen on the group is substituted by one or more substituents selected from the group consisting of: hydroxyl, methoxy, halogen, C1-C6 alkyl, C1-C6 alkoxy, carboxyl, sulfonic acid group, glucosamine group, morpholinyl, N-dimethyl.

4. The compound of claim 1, wherein R is₃And R₄Each independently is a C1-C4 alkyl group;

5. The compound of claim 1, wherein R is₆And R₇Each independently hydrogen, methyl or ethyl.

6. The compound of claim 1, wherein R is₈And R₉Each independently of the others is hydrogen, methyl, ethyl, n-propyl, isopropyl,

Wherein R is₁₂、R₁₃And R₁₄Each independently hydrogen, substituted or unsubstituted C1-C6 alkyl, R_cSO₂-, or R_cCO-wherein R_cIs hydrogen, hydroxyl, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 3-7 membered nitrogen-containing heterocyclic group, R_dNH-、R_dO(CH₂)_sNH-, or R_dO(CH₂)_s-; s is 0, 1, 2, 3 or 4, each R_dTaken from hydrogen, substituted or unsubstituted C1-C6 alkyl or substituted or unsubstituted 5-6 membered nitrogen containing heteroaryl;

b is NH or O, and B is NH or O,

R₁₅is hydrogen, substituted or unsubstituted C1-C6 alkyl, or 5-7 membered saturated heterocycle.

7. The compound of claim 1, wherein said compound is:

8. a pharmaceutical composition, comprising:

the compound of claim 1, a pharmaceutically acceptable salt thereof, or a dimer thereof; and

a pharmaceutically acceptable carrier.

9. The compound of claim 1 or the pharmaceutical composition for use of claim 8,

(i) for preparing bile acid G protein coupled receptor TGR5 agonist; or

(ii) Can be used for preparing medicine for treating metabolic diseases.

10. The use according to claim 9, wherein the metabolic disease is selected from the group consisting of: diabetes, obesity, hyperlipidemia, liver injury, and inflammatory diseases.