TWI395582B - Dicyclic azaalkane derivatives,preparing method and use thereof - Google Patents

Description

Azabicycloalkane derivative, preparation method thereof and use thereof in medicine

The present invention relates to an azabicycloalkane derivative, a process for the preparation thereof, a pharmaceutical composition containing the same, and its use as a therapeutic agent, in particular as a dipeptidyl peptidase inhibitor (DPPIV).

Diabetes is a multi-pathogenic metabolic disease characterized by chronic hyperglycemia accompanied by disorders of sugar, fat and protein metabolism caused by defects in insulin secretion and/or function. Diabetes is a very old disease caused by an increase in the concentration of glucose in the blood caused by absolute or relative lack of insulin in the human body. In turn, a large amount of sugar is excreted from the urine, and polydipsia, polyuria, polyphagia, weight loss, dizziness, Weakness and other symptoms.

Dipeptidyl peptidase-IV (DPPIV) is a serine protease that cleaves the N-terminal dipeptidase in the peptide chain containing a proline residue at the secondary end, despite the physiological effects of DPPIV on mammals. It has not been fully confirmed, but it plays an important role in neuroenzyme metabolism, T-cell initiation, cancer cell metastasis into the endothelium and HIV virus entry into lymphoid cells (WO 98/19998).

Recently, studies have shown that DPPIV can prevent the secretion of glucagon-like peptide (GLP)-1, in particular, it can cleave the N-terminal group-His-Ala dipeptidase in GLP-1 from The active form of GLP-1(7-36)NH ₂ degrades to inactive GLP-1(9-36)NH ₂ (Endocrinology, 1999, 140: 5356-5536). Due to physiological conditions, the half-life of intact GLP-1 in the blood of the loop is very short. The inactive metabolites after DPPIV degrades GLP-1 can bind to the GLP-1 receptor and antagonize the activity of GLP-1, thereby shortening the GLP. -1 physiological response. DPPIV inhibitors can completely protect endogenous and even exogenous GLP-1 from DPPIV deactivation, significantly increasing the physiological activity of GLP-1 (5 to 10 times), since GLP-1 is a secretory factor for pancreatic insulin secretion. Important stimulants can directly affect the distribution of glucose, and DPPIV inhibitors can play a good role in the treatment of non-insulin-dependent diabetes mellitus (NIDDM) (US6110949).

The present invention relates to a compound represented by the formula (I) or a pharmaceutically acceptable salt thereof:

Wherein: R is selected from the group consisting of an alkyl group, a cycloalkyl group, a halogenated alkyl group, an aryl group, a heteroaryl group, an amine alkyl group, a nonylamino group, a heterocyclic amine alkyl group or an amine alkyl group, wherein the heterocyclic ring is a 5-membered heterocyclic ring or a 6-membered heterocyclic ring, and the heterocyclic ring may be further subjected to one or more alkyl groups, aryl groups, heteroaryl groups, haloalkyl groups, haloalkoxy groups, hydroxyl groups, amine groups, alkylamino groups, decylamino groups. , an amine group, a cyano group, an alkynyl group, an alkoxy group, an aryloxy group, an amine alkyl group, a hydroxyalkyl group, a heterocycloalkyl group, a carboxyl group, a carboxylate group or a halogen; R _{1 is} selected from a hydrogen atom, an alkane a base, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, -C(O)NR ₃ R ₄ , -C(O)R ₃ , -C(O)OR ₃ , wherein the alkyl groups , cycloalkyl, heterocycloalkyl, aryl or heteroaryl may be further substituted by one or more alkyl, aryl, hydroxy, amine, alkoxy, aryloxy or heterocycloalkyl; R ₂ It is selected from a hydrogen atom or an alkyl group, wherein the alkyl group may be further substituted with one or more substituents selected from a cycloalkyl group or an aryl group; and R ₃ and R ₄ are each selected from a hydrogen atom, an alkyl group, and a cycloalkyl group; An aryl group, a heteroaryl group or a heterocycloalkyl group, wherein The alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl group may further be via one or more alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy Substituted, heteroaryloxy, halogen, hydroxy, amine, cyano, hydroxyalkyl, heterocycloalkyl, heterocycloalkoxy, trifluoromethyl, carboxy or carboxylate; or, R ₃ and R ₄ may form a 3 to 8 membered heterocyclic group together with the N atom, wherein the 5 to 8 membered heterocyclic ring may further contain one or more N, O or S atoms, and the 3 to 8 membered heterocyclic ring Further further via one or more alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkoxy, aryloxy, hydroxyalkyl, heterocycloalkyl, carboxy , a carboxylate group, a halogen or a -NR ₃ R ₄ substituent; n is 0 to 4.

Furthermore, the present invention includes a compound represented by the following formula (IA) or a pharmaceutically acceptable salt thereof:

Wherein: R is selected from the group consisting of an alkyl group, a cycloalkyl group, a halogenated alkyl group, an aryl group, a heteroaryl group, an amine alkyl group, a nonylamino group, a heterocyclic amine alkyl group or an amine alkyl group, wherein the heterocyclic ring is 5 a heterocyclic ring or a 6-membered heterocyclic ring, and the heterocyclic ring may further be subjected to one or more alkyl groups, aryl groups, heteroaryl groups, haloalkyl groups, haloalkoxy groups, hydroxyl groups, amine groups, alkylamino groups, decylamino groups, Amine, cyano, alkynyl, alkoxy, aryloxy, aminoalkyl, hydroxyalkyl, heterocycloalkyl, carboxy, carboxylate or halogen; R _{1 is} selected from a hydrogen atom, an alkyl group a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, -C(O)NR ₃ R ₄ , -C(O)R ₃ , -C(O)OR ₃ , wherein the alkyl group, cycloalkyl, heterocycloalkyl, aryl or heteroaryl groups may be further substituted with one or more alkyl, aryl, hydroxyl, amino, alkoxy, aryloxy, or substituted heterocycloalkyl; R ₂ is selected from a hydrogen atom or an alkyl group, wherein the alkyl group may be further substituted with one or more substituents selected from a cycloalkyl group or an aryl group; and R ₃ and R ₄ are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, An aryl group, a heteroaryl group or a heterocycloalkyl group, wherein Or a cycloalkyl, aryl, heteroaryl or heterocycloalkyl group may further be substituted by one or more alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy groups , heteroaryloxy, halogen, hydroxy, amine, cyano, hydroxyalkyl, heterocycloalkyl, heterocycloalkoxy, trifluoromethyl, carboxy or carboxylate; or, R ₃ and R ₄ may form a 3 to 8 membered heterocyclic group together with the N atom, wherein the 5 to 8 membered heterocyclic ring may further contain one or more N, O or S atoms, and the 3 to 8 membered heterocyclic ring may be Further by one or more alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkoxy, aryloxy, hydroxyalkyl, heterocycloalkyl, carboxy, Carboxylic acid ester group, halogen or -NR ₃ R ₄ substituted.

Preferably, in the compound of the formula (I) or a salt thereof, R is

Wherein R _{5 is} selected from a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group, a heteroaryl group or a heterocycloalkyl group, wherein the alkyl group, cycloalkyl group, aryl group, heteroaryl group or heterocycloalkyl group Further further via one or more alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy, heteroaryloxy, halogen, hydroxy, amine, alkylamine, Substituted with cyano, hydroxyalkyl, heterocycloalkyl, heterocycloalkoxy, carboxy or carboxylate; R ₆ and R ₇ are each selected from alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy , hydroxy, amine, cyano, alkynyl, alkoxy, aryloxy, hydroxyalkyl, heterocycloalkyl, carboxy, carboxylate or halogen; W is a carbon, sulfur or oxygen atom; when W is In the case of carbon, it may be further substituted with R ₆ or R ₇ .

Furthermore, the present invention includes the compound represented by the following (IB) or a pharmaceutically acceptable salt thereof:

Where: R is

R _{1 is} selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, -C(O)NR ₃ R ₄ , -C(O)R ₃ , -C(O)OR ₃ wherein the alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group may be further selected from one or more selected from the group consisting of an alkyl group, an aryl group, a hydroxyl group, an amine group, an alkoxy group, and an aryloxy group. Substituted with a substituent of a heterocycloalkyl group; R _{2 is} selected from a hydrogen atom or an alkyl group, wherein the alkyl group may be further substituted with one or more substituents selected from a cycloalkyl group or an aryl group; R ₃ and R _{4 is} each selected from a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group, a heteroaryl group or a heterocycloalkyl group, wherein the alkyl group, cycloalkyl group, aryl group, heteroaryl group or heterocycloalkyl group can be further One or more selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy, heteroaryloxy, halogen, hydroxy, amine, cyano, hydroxy Substituted by a substituent of an alkyl group, a heterocycloalkyl group, a heterocycloalkoxy group, a trifluoromethyl group, a carboxyl group or a carboxylate group; or, R ₃ and R ₄ together with the N atom form a 3 to 8 membered heterocyclic ring a base, wherein the 5 to 8 member heterocyclic ring may further contain one or more N, O or S atoms, and the 3 to 8 membered heterocyclic ring may further be selected from one or more selected from the group consisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkoxy Substituted with a substituent of a aryloxy group, an aryloxy group, a hydroxyalkyl group, a heterocycloalkyl group, a carboxyl group, a carboxylate group, a halogen or a -NR ₃ R ₄ ; R _{5 is} selected from a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group a heteroaryl or heterocycloalkyl group, wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl group may further be selected from one or more selected from the group consisting of alkyl, cycloalkyl, aryl , heteroaryl, alkoxy, cycloalkoxy, aryloxy, heteroaryloxy, halogen, hydroxy, amine, alkylamino, cyano, hydroxyalkyl, heterocycloalkyl, heterocycloalkoxy Substituted by a substituent of a carboxy group or a carboxylate group; R ₆ and R ₇ are each selected from alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkynyl, alkane Oxyl, aryloxy, hydroxyalkyl, heterocycloalkyl, carboxyl, carboxylate or halogen; W is a carbon, sulfur or oxygen atom; when W is carbon, it may be further substituted by R ₆ or R ₇ .

Further, the present invention includes a compound represented by the following formula (IC) or a pharmaceutically acceptable salt thereof:

Where: R is

R _{1 is} selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, -C(O)NR ₃ R ₄ , -C(O)R ₃ , -C(O)OR ₃ wherein the alkyl group, cycloalkyl group, heterocycloalkyl group, aryl group or heteroaryl group may be further selected from one or more selected from the group consisting of an alkyl group, an aryl group, a hydroxyl group, an amine group, an alkoxy group, and an aryloxy group. Substituted with a substituent of a heterocycloalkyl group; R _{2 is} selected from a hydrogen atom or an alkyl group, wherein the alkyl group may be further substituted with one or more substituents selected from a cycloalkyl group or an aryl group; R ₃ and R _{4 is} each selected from a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group, a heteroaryl group or a heterocycloalkyl group, wherein the alkyl group, cycloalkyl group, aryl group, heteroaryl group or heterocycloalkyl group can be further One or more selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy, heteroaryloxy, halogen, hydroxy, amine, cyano, hydroxy Substituted with a substituent of an alkyl group, a heterocycloalkyl group, a heterocycloalkoxy group, a trifluoromethyl group, a carboxyl group or a carboxylate group; or, R ₃ and R ₄ together with the N atom form a heterocyclic group of 3 to 8 members Wherein the 5 to 8 member heterocyclic ring may further comprise one or more N, O or S atoms, and the 3 to 8 membered heterocyclic ring may further be selected from one or more selected from the group consisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkoxy Substituted with a substituent of a aryloxy group, an aryloxy group, a hydroxyalkyl group, a heterocycloalkyl group, a carboxyl group, a carboxylate group, a halogen or a -NR ₃ R ₄ ; R _{5 is} selected from a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group a heteroaryl or heterocycloalkyl group, wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl group may further be selected from one or more selected from the group consisting of alkyl, cycloalkyl, aryl , heteroaryl, alkoxy, cycloalkoxy, aryloxy, heteroaryloxy, halogen, hydroxy, amine, alkylamino, cyano, hydroxyalkyl, heterocycloalkyl, heterocycloalkoxy Substituted by a substituent of a carboxy group or a carboxylate group; R ₆ and R ₇ are each selected from the group consisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkynyl, alkane Oxyl, aryloxy, hydroxyalkyl, heterocycloalkyl, carboxyl, carboxylate or halogen; W is a carbon, sulfur or oxygen atom; when W is carbon, it may be further substituted by R ₆ or R ₇ .

The pharmaceutically acceptable salt described in the present invention is a salt of the compound of the present invention and an acid selected from the group consisting of hydrochloric acid, p-toluenesulfonic acid, tartaric acid, maleic acid, lactic acid, methanesulfonic acid, sulfuric acid, phosphoric acid, and lemon. Acid, acetic acid or trifluoroacetic acid. Preference is given to p-toluenesulfonic acid, hydrochloric acid, tartaric acid or trifluoroacetic acid.

Specifically, the present invention includes compounds having the structures described below and salts thereof:

Further, the present invention relates to a compound represented by the following formula (ID) which is an intermediate for the synthesis of the compound of the formula (I) of the present invention:

Wherein: R _{1 is} selected from the group consisting of a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, -C(O)NR ₃ R ₄ , -C(O)R ₃ , -C(O And OR ₃ , wherein the alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may further be selected from one or more selected from the group consisting of an alkyl group, an aryl group, a hydroxyl group, an amine group, an alkoxy group, Substituted with a substituent of an aryloxy or heterocycloalkyl group; R ₃ and R ₄ are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group, a heteroaryl group or a heterocycloalkyl group, wherein the alkyl group, The cycloalkyl, aryl, heteroaryl or heterocycloalkyl group may further be selected from one or more selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy Substituted with a heteroaryloxy, halogen, hydroxy, amine, cyano, hydroxyalkyl, heterocycloalkyl, heterocycloalkoxy, trifluoromethyl, carboxy or carboxylate group; or, R ₃ and R ₄ together with the N atom form a 3 to 8 membered heterocyclic group, wherein the 5 to 8 membered heterocyclic ring may further contain one or more N, O or S atoms, and the 3 to 8 membered heterocyclic ring may further One or more selected from the group consisting of alkyl, aryl, heteroaryl, haloalkyl Substituted by haloalkoxy, hydroxy, amine, cyano, alkoxy, aryloxy, hydroxyalkyl, heterocycloalkyl, carboxy, carboxylate, halogen or substituents of -NR ₃ R ₄ .

The present invention relates to a process for the preparation of a compound of the formula (ID), comprising:

The 5-butoxy-perhydro-cyclopenta[c]pyrrole-2-carboxylic acid tert-butyl ester is reacted with trifluoroacetic acid in a suitable solvent (for example, dichloromethane) in an ice bath to obtain the whole Hydrogen-cyclopenta[c]pyrrole-5-one trifluoroacetate;

The perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate is reacted with hydrazine chloride or an ester in the presence of a base to give a compound of the formula (ID); the invention relates to the formula (IB) A method for preparing a compound, comprising:

The intermediate (ID) is reacted with a different amine, a substituted sodium borohydride, under basic conditions (such as triethylamine) in a suitable solvent (such as methanol, ethanol) at room temperature. A compound of the formula (IB) is obtained.

The present invention relates to a process for the preparation of a compound of the formula (IB), comprising:

After heating the toluene solution of potassium t-butoxide and methyltriphenylphosphonium iodide, the intermediate (ID) compound is added to the above solution at room temperature to obtain a methylene azabicyclo compound;

The methylene azabicyclo compound is reacted with trimethylsilyl cyanide in a suitable solvent (such as dichloromethane) in the presence of silver perchlorate at room temperature to give an isocyano aza Bicyclic compound;

The isocyanoazabicyclo compound is reacted in a suitable solvent (such as ethanol) with a suitable acid (such as hydrochloric acid) at room temperature to obtain an amine azabicyclo compound;

The amino azabicyclo compound is reacted with a halide in a solvent such as N,N-dimethylformamide to give a compound of the formula (IB).

The present invention relates to a process for the preparation of a compound of the formula (IB), comprising:

The intermediate (ID) compound and tosylmethylisonitrile are cyanated in a suitable solvent (such as ethylene glycol dimethyl ether) to obtain a cyano azabicyclo compound;

The cyanoazabicyclo compound is reacted with a halide in a suitable solvent (such as tetrahydrofuran) in the presence of lithium hexamethyldidecylamine to obtain an R ₂ -substituted cyanoazabicyclo compound;

The R ₂ -substituted cyanoazabicyclo compound is hydrolyzed by an acid to obtain an R ₂ -substituted carboxyazabicyclo compound; or the R ₂ -substituted cyanoazabicyclo compound is suitable. In a solvent such as dichloromethane, it is reacted with a reducing agent hydrogenated diisobutylaluminum (DIBAL-H) in an ice bath to first reduce the cyano group to an aldehyde group, and then the obtained aldehyde is mixed in a tetrahydrofuran/water mixed solvent. Reacting with sodium dihydrogen phosphate, sodium chlorite and 2-methyl-2-butene in an ice bath to obtain a R ₂ -substituted carboxyazabicyclo compound;

The R ₂ -substituted carboxyazabicyclo compound is reacted with an alkali azide (such as sodium azide) in the presence of basic conditions (such as triethylamine) and ethyl chloroformate to give an azide carbonyl group. Azabicyclo compound;

The azidocarbonylcarbonyl azabicyclo compound is heated in a suitable solvent (such as toluene), stirred in an acidic solution, and the pH is adjusted to be basic to obtain an R ₂ -substituted amino azabicyclo compound;

The R ₂ -substituted amino azabicyclo compound is reacted with a halide in a solvent such as N,N-dimethylformamide to give a compound of the formula (IB).

The present invention relates to a process for the preparation of a compound of the formula (IC), comprising:

The intermediate (ID) is reacted with a reducing agent (such as hydrogenated lithium trit-butoxide aluminum) in a suitable solvent (such as tetrahydrofuran) in an ice bath to obtain a hydroxyazabicyclo compound;

The hydroxyazabicyclo compound is reacted with an alkaline reagent (such as triethylamine) or methanesulfonyl chloride in a suitable solvent (such as dichloromethane) to obtain a methanesulfonyl azabicyclo compound;

The methanesulfonyl azabicyclo compound is heated in a solvent (such as N,N-dimethylformamide) with potassium phthalimide to obtain a phthalic acid imine substituted. Azabicyclo compound;

The phthalimide substituted azabicyclo compound is heated with hydrazine in a suitable solvent such as ethanol to obtain an amine azabicyclo compound;

The amine azabicyclo compound is heated and reacted with a halide in a suitable solvent such as dichloromethane to give a compound of the formula (IC).

Preferably, R is:

R _{5 is} selected from a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group, a heteroaryl group or a heterocycloalkyl group, wherein the alkyl group, cycloalkyl group, aryl group, heteroaryl group or heterocycloalkyl group may be further One or more selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy, heteroaryloxy, halogen, hydroxy, amine, alkylamine, Substituted with a substituent of a cyano group, a hydroxyalkyl group, a heterocycloalkyl group, a heterocycloalkoxy group, a carboxyl group or a carboxylate group; R ₆ and R ₇ are each selected from the group consisting of an alkyl group, an aryl group, a heteroaryl group, a halogenated alkyl group, Haloalkoxy, hydroxy, amine, cyano, alkynyl, alkoxy, aryloxy, hydroxyalkyl, heterocycloalkyl, carboxy, carboxylate or halogen; W is a carbon, sulfur or oxygen atom; When W is carbon, it may be further substituted with R ₆ or R ₇ .

Further, the present invention also includes the step of purifying the obtained compound of the formula (IB) or (IC) and reacting it with an acid in methanol, dichloromethane or ethyl acetate to obtain an acid addition product salt.

Further, the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

Furthermore, the present invention also relates to the use of a compound of the present invention or a pharmaceutically acceptable salt thereof for the preparation of a dipeptidyl peptidase (DPPIV) inhibitor drug.

In the compound of the formula (I) of the present invention or a salt thereof, the formula (I) is present in a free form or in an acid addition salt, and provides a pharmaceutically acceptable (non-toxic, physiologically acceptable) salt. Such pharmaceutically acceptable salts include hydrochloride, p-toluenesulfonate, tartrate, maleate, lactate, Sulfonate, sulfate, phosphate, citrate, acetate or trifluoroacetate. Preferred are p-toluenesulfonate, hydrochloride, tartrate and trifluoroacetate.

Unless otherwise stated, the following terms used in the specification and claims have the following meanings.

"Alkyl" means a saturated aliphatic hydrocarbon group comprising straight and branched chain groups of 1 to 20 carbon atoms. A medium-sized alkyl group having 1 to 10 carbon atoms is preferred, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, pentyl and the like. More preferably, it is a lower alkyl group having 1 to 4 carbon atoms, such as a methyl group, an ethyl group, a propyl group, a 2-propyl group, a n-butyl group, an isobutyl group or a t-butyl group. The alkyl group may be substituted or unsubstituted, and when substituted, preferred substituents are halogen, hydroxy, lower alkoxy, aryl, aryloxy, heteroaryl, heterocycloalkyl, C(O). R ₃ and C(O)NR ₃ R ₄ .

"Cycloalkyl" means a 3 to 8 membered all-carbon monocyclic, all-carbon 5/6 or 6/6 member fused or polycyclic fused ring ("fused" ring means each in the system The rings share an adjacent pair of carbon atoms) groups with other rings in the system, wherein one or more of the rings may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system. Examples of cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cyclopentene, cyclohexane, cyclohexadiene, adamantane, cycloheptane, cycloheptatriene and the like. The cycloalkyl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more substituents independently selected from the group consisting of lower alkyl, trihaloalkyl, halogen, hydroxy, lower alkoxy, aryl (via one or more Substituted independently of a halogen, hydroxy, lower alkyl or lower alkoxy group, aryloxy (which may be substituted by one or more substituents independently selected from halo, hydroxy, lower alkyl or lower alkoxy) Substituted), 6 membered heteroaryl (having 1 to 3 nitrogen atoms in the ring, the carbon in the ring may be substituted by one or more groups as desired, and the substituents are independently of each other halogen, hydroxy, lower alkyl or Lower alkoxy), 5-membered heteroaryl (having 1 to 3 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, the carbon and nitrogen atoms of which may optionally be selected from halogen, by one or more a substituent of a hydroxy group, a lower alkyl group or a lower alkoxy group), or a 5- or 6-membered heterocycloalkyl group (having 1 to 3 hetero atoms selected from nitrogen, oxygen and sulfur, carbon and nitrogen atoms of the group) (if any) may be substituted, if desired, with one or more substituents independently selected from halo, hydroxy, lower alkyl or lower alkoxy) , mercapto, (lower alkyl)thio, arylthio (optionally substituted with one or more substituents independently selected from halogen, hydroxy, lower alkyl or lower alkoxy), cyano, fluorenyl , thiodecyl, O-aminomethylindenyl, N-aminomethylindenyl, O-thioaminomethylindenyl, N-thioaminomethylindenyl, C-decylamine, N- Amidino, nitro, N-sulfonylamino, S-sulfonylamino, C(O)R ₃ , C(O)NR ₃ R ₄ and -C(O)OR ₃ .

"Alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond. Representative examples include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, and the like.

"Alkynyl" means an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond. Representative examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, and the like.

"Aryl" means a group having at least one aromatic ring structure, ie having a total The aromatic ring of the π-electron system of the yoke includes a carbocyclic aryl group, a heteroaryl group, and a biaryl group. The aryl group may be optionally substituted with one or more of the following substituents: halogen, trihalomethyl, hydroxy, SR, nitro, cyano, alkoxy and alkyl.

"Heteroaryl" refers to an aryl group having from 1 to 3 heteroatoms as ring atoms, the remaining ring atoms being carbon, and heteroatoms including oxygen, sulfur and nitrogen. The ring can be a 5 or 6 member ring. Examples of the heterocyclic aryl group include a furyl group, a thienyl group, a pyridyl group, a pyrrolyl group, an N-alkylpyrrolyl group, a pyrimidinyl group, a pyrazinyl group, an imidazolyl group and the like.

"Heterocycloalkyl" means a monocyclic or fused ring radical having from 5 to 9 ring atoms in the ring wherein one or two ring atoms are selected from nitrogen, oxygen or S(O)n (where n is an integer) From 0 to 2), the remaining atoms are carbon. These rings may also have one or more double bonds, however, these rings do not have a fully conjugated pi-electron system. Unsubstituted heterocycloalkyl groups include, but are not limited to, pyrrolidinyl, piperidin-1-yl, piperazin-1-yl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc., heterocycloalkyl It may be substituted or unsubstituted. When substituted, the substituent is preferably one or more, more preferably one, two or three, and even more preferably one or two, independently selected from the group consisting of lower alkyl, trihaloalkyl, halogen, hydroxy, A group consisting of lower alkoxy groups, cyano groups and fluorenyl groups. Preferably, the heteroaryl group may be substituted with one or two substituents independently selected from halo, lower alkyl, trihaloalkyl, hydroxy, decyl, cyano, N-nonylamino or carboxy.

"Hydroxy" refers to an -OH group.

"Alkoxy" means -O-(alkyl) and -O-(unsubstituted cycloalkyl). Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, and the like.

"Haloalkoxy" means -O-(haloalkyl). Representative examples include, but are not limited to, trifluoromethoxy, tribromomethoxy, and the like.

"Aryloxy" means -O-aryl and -O-heteroaryl, and aryl and heteroaryl are as defined above. Representative examples include, but are not limited to, phenoxy, pyridyloxy, furanoxy, thienyloxy, pyrimidinyloxy, pyrazinyloxy, and the like, and derivatives thereof.

"Amidino" refers to a -SH group.

"Alkylthio" means -O-(alkyl) and -O-(unsubstituted cycloalkyl). Representative examples include, but are not limited to, methylthio, ethylthio, propylthio, butylthio, cyclopropylthio, cyclobutylthio, cyclopentylthio, cyclohexylthio, and the like.

"Arylthio" means -S-aryl and -S-heteroaryl, and aryl and heteroaryl are as defined above. Representative examples include, but are not limited to, phenylthio, pyridylthio, furanthio, thiophenothionyl, pyrimidinyl, and the like, and derivatives thereof, "mercapto" refers to a -C(O)-R" group, wherein R "selected from hydrogen, lower alkyl, trihalomethyl, unsubstituted cycloalkyl, aryl (optionally substituted by one or more, preferably one, two or three substituents, the substituent) Selected from the group consisting of lower alkyl, trihalomethyl, lower alkoxy and halogen), heteroaryl (bonded by ring carbon) (optionally one or more, preferably one, two or three) Substituted by a substituent selected from the group consisting of lower alkyl, trihalomethyl, lower alkoxy and halogen) and heteroalicyclic (via ring carbon bonding) (optionally one or more Preferably, one, two or three substituents are selected which are selected from the group consisting of lower alkyl, trihalomethyl, lower alkoxy and halogen). Representative sulfhydryl groups include, but are not limited to, ethenyl, trifluoroethenyl, benzhydryl, and the like.

"Thiodecyl" refers to a -C(S)-R" group wherein R" is as defined above.

"Acetyl" group refers to a -C (O) CH ₃ group.

"Halogen" means fluoro, chloro, bromo or iodo, preferably fluoro or chloro.

"Trifluoromethyl" means -CF ₃ .

"Cyano" means -CN.

"Amino" means -NH _2.

"Carboxylic acid" means -COOH.

"Carboxylic acid ester" means -COOR wherein R is alkyl or cycloalkyl.

"Hydroxyalkyl" means -(CH2)rNH2 wherein r is from 1 to 4.

"As needed" or "as needed" means that the subsequently described event or environment may, but need not, occur, including where the event or environment occurs or does not occur. For example, "heterocyclic group which may be optionally substituted by an alkyl group" means that an alkyl group may exist but is not necessarily present, and the description includes the case where the heterocyclic group is substituted with an alkyl group and the heterocyclic group is not substituted with an alkyl group. The situation.

"Pharmaceutical composition" means a mixture of one or more of the compounds described herein or a physiologically/pharmaceutically acceptable salt or prodrug thereof with other chemical components, such as physiologically/pharmaceutically acceptable carriers. And excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.

Method for synthesizing the compound of the present invention

In order to accomplish the object of the present invention, the present invention adopts the following technical solutions:

Synthetic scheme I

5-butyloxy-perhydro-cyclopenta[c]pyrrole-2-carboxylic acid tert-butyl ester is reacted with trifluoroacetic acid in a suitable solvent (such as dichloromethane) in an ice bath to obtain the whole Hydrogen-cyclopenta[c]pyrrole-5-one trifluoroacetate; reacting perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate with hydrazine or an ester in the presence of a base a compound of the formula (ID); a different amine, substituted sodium borohydride of the intermediate (ID) in a suitable solvent (such as methanol, ethanol) at room temperature under basic conditions (eg Triethylamine) is reacted to give a compound of the formula (IB).

Synthetic Scheme II

After heating the toluene solution of potassium t-butoxide and methyltriphenylphosphonium iodide, the intermediate (ID) compound is added to the above solution at room temperature to obtain a methylene azabicyclo compound; The methylene azabicyclo compound is reacted with trimethylocyanine at room temperature in a suitable solvent (such as dichloromethane) in the presence of silver perchlorate to give an isocyanoazabicyclo compound; The isocyanoazabicyclo compound is reacted in a suitable solvent (such as ethanol) with a suitable acid (such as hydrochloric acid) at room temperature to obtain an amine azabicyclo compound; the amine azabicyclo compound is in a solvent (eg N,N-dimethyl A The reaction with a halide in a solvent gives the compound of the formula (IB).

Synthetic scheme III

The intermediate (ID) compound and p-toluenesulfonylmethyl isocyanide are cyanated in a suitable solvent (such as ethylene glycol dimethyl ether) to obtain an azabicyclo compound; the cyano azabicyclo compound is Reacting with a halide in the presence of lithium hexamethyldidecylamine in a suitable solvent (such as tetrahydrofuran) to give an R ₂ -substituted cyanoazabicyclo compound; the R ₂ -substituted azabicyclo ring The compound is hydrolyzed by an acid to give an R ₂ -substituted carboxyazabicyclo compound, or the R ₂ -substituted cyanoazabicyclo compound is dissolved in a suitable solvent (such as dichloromethane) in an ice bath. The reducing agent hydrogenated diisobutylaluminum (DIBAL-H) reaction, first reducing the cyano group to an aldehyde group, and then the obtained aldehyde in a tetrahydrofuran / water mixed solvent, in an ice bath with sodium dihydrogen phosphate, chlorous acid Reaction of sodium with 2-methyl-2-butene to give a R ₂ -substituted carboxyazabicyclo compound; the R ₂ -substituted carboxyazabicyclo compound under basic conditions (such as triethylamine) and chlorine In the presence of ethyl formate, reacting with an azide alkali metal such as sodium azide An azidocarbonylcarbonyl azabicyclo compound; heating the azidocarbonyl azabicyclo compound in a suitable solvent (such as toluene), stirring in an acidic solution, and adjusting the pH to be basic to obtain an R ₂ -substituted An amino azabicyclo compound; reacting the R ₂ -substituted amino azabicyclo compound in a solvent (such as N,N-dimethylformamide) with a halide to give a compound of the formula (IB).

Synthetic Scheme IV

The intermediate (ID) is reacted with a reducing agent (such as hydrogenated lithium trit-butoxide aluminum) in a suitable solvent (such as tetrahydrofuran) to obtain a hydroxyazabicyclo compound; the hydroxyazabicyclo compound is In a suitable solvent (such as dichloromethane), react with an alkaline reagent (such as triethylamine) or methanesulfonyl chloride in an ice bath to obtain a methanesulfonyl azabicyclo compound; the methanesulfonyl azabicyclo ring The compound is heated in a solvent (such as N,N-dimethylformamide) and potassium phthalimide to obtain an azabibiamine substituted with phthalimide; the orthobenzene The dimethyl quinone substituted azabicyclo compound is heated with hydrazine in a suitable solvent (such as ethanol) to give an amine azabicyclo compound; the amine azabicyclo compound is in a suitable solvent (eg dichloro Methane), heated with a halide, is reacted to give a compound of the formula (IC).

Further, after purifying the compound of the formula (IB) or (IC), it is reacted with an acid in methanol, dichloromethane or ethyl acetate to obtain an acid addition product salt.

The present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound or a salt thereof and a pharmaceutical carrier, or a compound of the formula (I) of the present invention or The use of a salt for the preparation of a dipeptidyl peptidase inhibitor drug. In other words, the invention also provides compositions comprising the above-described compounds in a pharmaceutically effective amount, and the use of such compounds in the preparation of dipeptidyl peptidase inhibitors.

detailed description

The invention is further described by the following examples, which are not intended to limit the scope of the invention.

Example

The structure of the compound is determined by nuclear magnetic resonance ( ¹ H NMR) or mass spectrometry (MS). ^{The 1} H NMR shift (δ) is expressed in parts per million (ppm). The NMR was measured by a Bruker AVANCE-400 nuclear magnetic apparatus, and the solvent was deuterated chloroform (CDCl ₃ ), deuterated dimethyl hydrazine (DMSO-D ₆ ), deuterated methanol (CD ₃ OD), and the internal standard was four. Methyl decane (TMS), chemical shift is expressed in units of 10 ^-6 (ppm).

The MS was measured using a FINNIGAN LCQAd (ESI) mass spectrometer.

Determination of average kinase inhibition rate and IC ₅₀ value using NovoStar microplate reader (BMG, Germany)

Thin layer of silicone rubber using Yantai Yellow Sea HSGF254 or Qingdao GF254 rubber sheet

Pipe column chromatography generally uses Yantai Huanghai Tanji 200~300 mesh silicone as carrier.

DMSO-D ₆ : deuterated dimethyl hydrazine; CDCl ₃ : deuterated chloroform; CD ₃ OD: deuterated methanol;

Example 1

Cis-5-(2-(S)-(2-cyanopyrrolidin-1-yl)-2-oxoethoxyethylamino)-N,N-dimethyl-perhydro-cyclopenta[c Pyrrole-2-carboxamide hydrochloride

first step

Preparation of [2-(2-aminomethylindenyl-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamic acid tert-butyl ester 1b

N-Tertioxycarbonyl-glycine 1a (5 g, 28.56 mmol) and L-amine amine (3.25 g, 28.50 mmol) were dissolved in 75 mL of N,N-dimethylformamide. , cooled to 0 ° C, and added 1-hydroxybenzotriazole (11.8 g, 87.3 mmol), N-ethyl-N'-(dimethylaminopropyl)-carbodiimide (11.3 g, 59) with stirring. Methyl) (12.1 mL, 87.3 mmol), naturally warmed to room temperature, stirred overnight, and the next day was followed by thin layer chromatography to trace the reaction until the material disappeared completely, and the N, N-di was evaporated at less than 50 ° C. Methyl carbamide, the reaction mixture was extracted with ethyl acetate (200 mL×3), and the organic phase was combined and washed with 50 mL of saturated sodium chloride. Recrystallization from ethyl acetate gave the title product [2-(2-aminocarbazin-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamic acid tert-butyl ester 1b (7.42 g, white powder), yield 95.8%.

MS m/z (ESI): 2721. (M + 1).

Second step

Preparation of [2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamic acid tert-butyl ester 1c

Under a nitrogen atmosphere, 286 mL of pyridine was added to a dry three-necked flask, [2-(2-aminoformamidine-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamic acid Tributyl ester 1b (13.5 g, 49.8 mmol), imidazole (7.11 g, 104.6 mmol), the reaction system was cooled to -35 ° C, and phosphorus oxychloride (19 mL, 204.2 mmol) was added dropwise with stirring at this temperature. The reaction was continued for 1 hour, then naturally warmed to room temperature. After 0.5 hours of reaction, the pyridine was evaporated to dryness, diluted with ethyl acetate, water was added, and the mixture was extracted with ethyl acetate (200 mL×3). The residue was washed with EtOAc (EtOAc m.) 1-Bis-2-yloxy-ethyl]-carbamic acid tert-butyl ester 1c (10.7 g, white powder), yield 84.9%.

MS m/z (ESI): 254.3 (M + 1).

third step

Preparation of 1-(2-aminoethenyl)-2-cyano-pyrrolidine hydrochloride 1d

[2-(2-Cyano-pyrrolidin-1-yl)-2-yloxy-ethyl]-carbamic acid tert-butyl ester 1c (13.7 g, 54.2 mmol) was dissolved in 140 mL of diethyl ether with stirring. And 40 mL of water, 37% hydrochloric acid solution (90 mL) was added dropwise in an ice bath, and the reaction was carried out for 1 hour in an ice bath, and the solvent was evaporated to dryness, and the mixture was evaporated to give the title product 1-(2-aminoethyl hydrazyl). 2-cyano-pyrrolidine hydrochloride 1d (10 g, white powder), yield 98%.

MS m/z (ESI): 154.4 (M + 1).

the fourth step

Preparation of perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate 1f

5-tert-oxy-perhydro-cyclopenta[c]pyrrole-2-carboxylic acid tert-butyl ester 1e (0.32 g, 1.42 mmol) was dissolved in 10 mL of dichloromethane under stirring, and added to the ice bath. trifluoroacetic acid (3.27 mL, 42.7 mmol), at 0 ℃ 30 minutes, the solvent was evaporated under reduced pressure to give the title product perhydro - cyclopenta [c] pyrrol-5-one trifluoroacetate 1f the crude product, which Used directly in the next step.

MS m/z (ESI): 126.4 (M + 1).

the fifth step

Preparation of N,N-Dimethyl-5-oxo-perhydro-cyclopenta[c]pyrrole-2-carboxamide 1g

The crude product of the previous step of perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate 1f was dissolved in 15 mL of acetonitrile with stirring, and potassium carbonate (0.24 g, 1.71 mmol) was added thereto under ice cooling. N,N-dimethylamine formazan chloride (0.14 mL, 1.56 mmol) was added dropwise, and the mixture was reacted for 2 hr at room temperature. The solvent was evaporated, 50 mL water was added, and ethyl acetate (50 mL×3) The organic phase was washed with 50 mL of EtOAc EtOAc. Dimethyl-5-oxo-perhydro-cyclopenta[c]pyrrole-2-carboxamide 1 g (0.19 g, pale yellow oil), yield 68.3%.

MS m/z (ESI): 197.4 (M + 1).

_{^{1 H NMR (DMSO-D 6}} , 400MHz) δ 3.56 (m, 2H), 2.85 (m, 2H), 2.7 (s, 6H), 2.81 (m, 2H), 2.5 (m, 2H), 2.01 (m , 2H).

Step 6

Cis-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2 - Preparation of methotrexate 1h

1-(2-Aminoethyl fluorenyl)-2-cyano-pyrrolidine hydrochloride 1d (0.36 g, 1.91 mmol) was dissolved in 20 mL of methanol with stirring, and N,N-dimethyl-5- was added. 1-oxy-perhydro-cyclopenta[c]pyrrole-2-carboxamide 1g (0.25 g, 1.28 mmol) and sodium triethoxysulfonylborohydride (1.22 g, 5.74 mmol), 3 at room temperature The reaction solution was concentrated, extracted with 20 mL of saturated sodium carbonate solution and then dichloromethane (20 mL×10). The organic phase was combined and washed with 10 mL of saturated sodium chloride. The organic layer was dried (MgSO4), filtered Oxyethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 1 h (0.3 g, white powder), yield 53%.

MS m/z (ESI): 334.5 (M + 1).

Seventh step

Cis-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2 -Preparation of methotrexate hydrochloride 1

Cis-5-(2-(2-cyanopyrrolidin-1-yl)-2-oneoxyethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole- 2-Methylamine 1h (200 mg, 0.687 mmol) was dispersed in 10 mL of dichloromethane, and 2 mL of 0.5N hydrochloric acid diethyl ether solution was added to an ice bath, and the solvent was evaporated to dryness. The product cis-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole- 2-Metamine hydrochloride 1 (180 mg, white powder), yield 80%.

¹ H NMR (CD ₃ OD, 400 MHz) δ 4.82 (dd, 1H, J ₁ = 4 Hz, J ₂ = 5.2 Hz), 4.02 (dd, 2H, J ₁ = J ₂ = 16.4 Hz), 3.62-3.25 (m) , 7H), 2.76 (s, 6H), 2.51-1.49 (m, 10H).

Example 2

Cis-5-(2-(S)-(2-cyanopyrrolidin-1-yl)-2-oxoethoxyethylamino)-perhydro-cyclopenta[c]pyrrole-2-carboxylic acid Ester hydrochloride

first step

Preparation of 5-sided oxy-perhydro-cyclopenta[c]pyrrole-2-carboxylic acid methyl ester 2a

The perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate 1f (0.559 g, 2.34 mmol) was dissolved in 20 mL of acetonitrile with stirring, and potassium carbonate (0.646 g, 4.68) was added to the ice water bath. Methyl chloroformate (0.22 mL, 2.8 mmol), mp EtOAc (EtOAc: EtOAc) After washing with 50 mL of a saturated sodium chloride solution and 50 mL of water, the ethyl acetate phase was dried over anhydrous magnesium sulfate and filtered, and the filtrate was concentrated under reduced pressure. -Sideoxy-perhydro-cyclopenta[c]pyrrole-2-carboxylic acid methyl ester 2a (0.25 g, colorless oil), yield 58.4%.

MS m/z (ESI): 184 (MH).

Second step

Preparation of methyl cis-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-perhydro-cyclopenta[c]pyrrole-2-carboxylate 2b

1-(2-Aminoethyl fluorenyl)-2-cyano-pyrrolidine hydrochloride 1d (0.43 g, 2.29 mmol) was dissolved in 20 mL of methanol with stirring. Methyl cyclopenta[c]pyrrole-2-carboxylate 2a (0.28 g, 1.53 mmol) and sodium triethoxysulfonylborohydride (1.46 g, 6.88 mmol) were reacted at room temperature for 3 hr. The reaction mixture was extracted with 20 mL of saturated sodium carbonate solution and then dichloromethane (20 mL×3). The organic phase was combined and washed with 10 mL of saturated sodium chloride. Concentration under reduced pressure and purification by silica gel column chromatography to give the product cis-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoxyethylamine)-perhydro-cyclopentane And [c]methyl pyrrole-2-carboxylate 2b (0.22 g, white powder), yield 41%.

MS m/z (ESI): 357 (MH).

third step

Cis-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-perhydro-cyclopenta[c]pyrrole-2-carboxylic acid methyl ester hydrochloride Preparation of 2

Disperse cis-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-perhydro-cyclopenta[c]pyrrole-2-carboxylic acid methyl ester 2b 2 mL of 0.5 N hydrochloric acid diethyl ether solution was added to 10 mL of diethyl ether under ice-cooling, and the precipitate was centrifuged to obtain the title product cis-5-(2-((S)-2-cyanopyrrolidin-1-yl). 2-oxoethoxyethylamino)-perhydro-cyclopenta[c]pyrrole-2-carboxylic acid methyl ester hydrochloride 2 (200 mg, white powder).

¹ H NMR (CD ₃ OD, 400 MHz) δ 4.71 (m, 1H), 3.93 (m, 2H), 3.59-3.28 (m, 10H), 2.64 (m, 2H), 2.34 (m, 2H), 2.17 ( m, 2H), 2.08 (m, 2H).

Example 3

Cis-1-(2-(2-(2-hydroxyethyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethenyl)-2-cyano-pyrrolidine hydrochloride salt

first step

Preparation of 2-(2-hydroxyethyl)-perhydro-cyclopenta[c]pyrrole-2-one 3a

All hydrogen-cyclopenta[c]pyrrole-5-one trifluoroacetate 1f (764.8 mg, 3.2 mmol) and 2-hydroxyacetic acid (267.5 mg, 3.52 mmol) were dissolved in 10 mL of acetonitrile with stirring, ice-water bath Glycolic acid (1.3 g, 9.6 mmol) and 1-ethyl-3-dimethylaminopropyl-carbodiimide hydrochloride (1.23 g, 6.4 mmol) and triethylamine (1.3 mL, 9.6 mmol) The ice water bath was removed, and the reaction was carried out at 25 ° C overnight. The solvent was evaporated to dryness. EtOAc was evaporated. concentrated and the resulting residue was purified by silica gel column chromatography to obtain the title product 2- (2-acetyl-yl) - perhydro - cyclopenta [c] pyrrol-2-one 3a (0.375 g, colorless oil ()), yield 64%.

MS m/z (ESI): 184 (MH).

Second step

Cis-1-(2-(2-(2-hydroxyethyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano-pyrrolidine 3b preparation

2-(2-Hydroxyethyl)-perhydro-cyclopenta[c]pyrrole-2-one 3a (0.375 g, 2.05 mmol) and 1-(2-aminoethenyl)-2-cyanide The pyridine-pyrrolidine hydrochloride 1d (0.78 g, 4.1 mmol) was dissolved in 10 mL of tetrahydrofuran and 5 mL of methanol with stirring, and reacted at room temperature for 0.5 hour, then sodium triethyloxyborohydride (0.87 g, 4.1) was added. The reaction mixture was stirred at rt. EtOAc was evaporated. Purification to give the product cis-1-(2-(2-(2-hydroxyethyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano Pyrrolidine 3 which was used directly in the next reaction.

MS m/z (ESI): 357 (MH).

third step

Cis-1-(2-(2-(2-hydroxyethyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethenyl)-2-cyano-pyrrolidine hydrochloride Preparation of salt 3

Cis-1-(2-(2-(2-hydroxyethyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano-pyrrolidine 3b Disperse in 10 mL of diethyl ether, add 2 mL of 0.5 N hydrochloric acid diethyl ether solution under ice bath, and centrifuge the precipitate to obtain the title product cis-1-(2-(2-(2-hydroxyethyl)-perhydro-ring Pentyl[c]pyrrole-5-ylamino)ethynyl)-2-cyano-pyrrolidine hydrochloride 3 (100 mg, white powder).

Example 4

Cis-1-(2-(2-(piperidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethenyl)-2-cyano-pyrrolidine hydrochloride salt

first step

Preparation of 2-(piperidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-one 4a

Dissolve perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate 1f (478 mg, 2 mmol) in 20 mL of dichloromethane and add iodination [3-(1-piperidine) Indole-based imidazol-1-methyl] (0.96 g, 3 mmol) and triethylamine (0.84 mL, 6 mmol), allowed to react at room temperature overnight, then quenched with 20 mL of water, with dichloromethane (50 mL) ×3) Extraction, the organic phase was combined, washed successively with 50 mL of 10% citric acid solution and 50 mL of saturated sodium chloride solution, and the dichloromethane phase was dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by chromatography to give the title product 2- (piperidin-1-carbonyl) - perhydro - cyclopenta [c] pyrrol-5-one 4a (0.41 g, colorless oil), yield 87 %.

MS m/z (ESI): 237 (MH).

Second step

Cis-1-(2-(2-(piperidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano-pyrrolidine 4b preparation

2-(Piperidine-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-one 4a (0.41 g, 1.74 mmol) and 1-(2-aminoethenyl)-2-cyanide The pyridine-pyrrolidine hydrochloride 1d (0.5 g, 2.6 mmol) was dissolved in 50 mL of tetrahydrofuran, and 5 g of sodium sulfate and 0.05 mL of acetic acid were added thereto, and the reaction was carried out at room temperature for 0.5 hour, followed by the addition of sodium triethoxysulfonate. 1.1 g, 5.2 mmol), after reacting for 3 hours at room temperature, the reaction mixture was concentrated under reduced pressure, then 50 mL of saturated sodium carbonate solution was added and extracted with ethyl acetate (50 mL×3). The sodium chloride solution and 50 mL of water were washed, and the ethyl acetate layer was dried over anhydrous magnesium sulfate. (Piperidine-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethynyl)-2-cyano-pyrrolidine 4b which was used directly in the next reaction.

MS m/z (ESI): 410 (MH).

third step

Cis-1-(2-(2-(piperidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethenyl)-2-cyano-pyrrolidine hydrochloride Preparation of salt 4

Cis-1-(2-(2-(piperidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano-pyrrolidine 4b Disperse in 10 mL of diethyl ether, add 2 mL of 0.5 N hydrochloric acid diethyl ether solution under ice bath, and centrifuge the precipitate to obtain the title product cis-1-(2-(2-(piperidin-1-carbonyl)-perhydro- Cyclopenta[c]pyrrole-5-ylamino)ethynyl)-2-cyano-pyrrolidine hydrochloride 4 (0.16 g, white powder).

¹ H NMR (CD ₃ OD, 400 MHz) δ 4.83 (dd, 1H, J ₁ = 3.0 Hz, J ₂ = 5.8 Hz), 4.09 (dd, 2H, J ₁ = J ₂ = 13.1 Hz), 3.70 - 3.30 ( m, 10H), 2.72 (m, 2H), 2.47 (m, 2H), 2.31-2.00 (m, 5H), 1.66-1.52 (m, 8H).

Example 5

Cis-1-(2-(2-ethylindenyl-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano-pyrrolidine hydrochloride

first step

Preparation of 2-ethylindenyl-perhydro-cyclopenta[c]pyrrole-5-one 5a

The perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate salt 1f (717 mg, 3 mmol) was dissolved in acetonitrile (20 mL) with stirring, and dibutyl succinate was added in an ice water bath ( 0.42 mL, 4.5 mmol) and triethylamine (0.98 mL, 9 mmol), EtOAc (EtOAc) EtOAc (EtOAc) The residue was washed with 50 mL of EtOAc EtOAc. [c] Pyrrol-5-one 5a (0.36 g, colorless oil), yield 72%.

MS m/z (ESI): 168.4 (M + 1).

Second step

Preparation of cis-1-(2-(2-ethylindenyl-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano-pyrrolidine 5b

2-Ethyl-perhydro-cyclopenta[c]pyrrole-5-one 5a (0.36 g, 2.15 mmol) and 1-(2-aminoethenyl)-2-cyano-pyrrolidine salt The acid salt 1d (0.614 g, 3.23 mmol) was dissolved in 50 mL of tetrahydrofuran, and 5 g of sodium sulfate and 0.05 mL of acetic acid were added. After reacting for 0.5 hour at room temperature, sodium triethoxysulfonate hydride (1.37 g, 6.46 mmol) was added. After reacting for 3 hours at room temperature, the reaction solution was concentrated under reduced pressure, and 50 mL of saturated sodium carbonate solution was added, and extracted with ethyl acetate (50 mL×3), and the organic phase was combined with 50 mL of saturated sodium chloride solution and 50 The title product was cis-1-(2-(2-ethenyl-perhydro). The titled product was purified by EtOAc EtOAc. -Cyclopenta[c]pyrrole-5-ylamino)ethynyl)-2-cyano-pyrrolidine 5b which was used directly in the next reaction.

MS m/z (ESI): 305.5 (M + 1).

third step

Preparation of cis-1-(2-(2-ethylindenyl-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano-pyrrolidine hydrochloride 5

The cis-1-(2-(2-ethylindenyl-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano-pyrrolidine 5b obtained in the previous step was dispersed in In 20 mL of diethyl ether, 4 mL of 0.5 N hydrochloric acid diethyl ether solution was added to an ice bath, and the precipitate was centrifuged to obtain the title product cis-1-(2-(2-ethyl)-perhydro-cyclopenta[c]pyrrole- 5-Aminoamino)ethinyl)-2-cyano-pyrrolidine 5 (0.23 g, white powder).

¹ H NMR (CD ₃ OD, 400 MHz) δ 4.71 (m, 1H), 3.92 (m, 2H), 3.69-3.37 (m, 7H), 2.69 (m, 2H), 2.33 (m, 2H), 2.13 ( m, 2H), 2.04-2.00 (m, 5H), 1.48 (m, 2H).

Example 6

Cis-5-[2-(2-cyanopyrrolidin-1-yl)-2-yloxy-ethylamino]-N-isopropylperhydro-cyclopenta[c]pyrrole-2-yl Indoleamine hydrochloride

first step

Preparation of N-isopropyl-5-sideoxy-perhydro-cyclopenta[c]pyrrole-2-carboxamide 6a

Dissolve perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate 1f (717 mg, 3 mmol) in 20 mL of dichloromethane with stirring and add isopropyl isocyanate (9 mL). , 9 mmol) and triethylamine (1.7 mL, 12 mmol). After reacting at room temperature overnight, add 50 mL of water, extract with dichloromethane (50 mL×3), combine the organic phases, and wash with 50 mL of 10% citric acid solution and 50 mL of saturated sodium chloride solution, respectively. The organic layer was dried over anhydrous magnesium sulfate, filtered, evaporated, evaporated, evaporated,,,,,,,,,,,, 2-carbamamine 6a (0.3 g, colorless oil), yield 47.6%

MS m/z (ESI): 211 (M + 1).

Second step

Cis-5-[2-(2-cyanopyrrolidin-1-yl)-2-yloxy-ethylamino]-N-isopropyl-perhydro-cyclopenta[c]pyrrole-2- Preparation of formamide 6b

N-Isopropyl-5-sideoxy-perhydro-cyclopenta[c]pyrrole-2-carboxamide 6a (0.3 g, 1.43 mmol) and (S)-1-(2-amino B Hydrazinyl-2-cyano-pyrrolidine hydrochloride 1d (0.407 g, 2.14 mmol) was dissolved in 50 mL of tetrahydrofuran, 5 g of sodium sulfate and 0.05 mL of acetic acid were added, and reacted at room temperature for 0.5 hour, then added with triethyl Sodium bis oxyborohydride (0.9 g, 4.3 mmol) was reacted at room temperature for 3 hr. The reaction mixture was evaporated. mjjjjjjjjjjj The phase was washed with 50 mL of a saturated sodium chloride solution and 50 mL of water, and the ethyl acetate layer was dried over anhydrous magnesium sulfate. -[2-(2-cyanopyrrolidin-1-yl)-2-oxo-ethylamino]-N-isopropyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 6b , which is used directly for the next reaction.

MS m/z (ESI): 384 (MH).

third step

Cis-5-[2-(2-cyanopyrrolidin-1-yl)-2-yloxy-ethylamino]-N-isopropyl-perhydro-cyclopenta[c]pyrrole-2- Preparation of formamide hydrochloride 6

The cis-5-[2-(2-cyanopyrrolidin-1-yl)-2-oxo-ethylamino]-N-isopropyl-perhydro-cyclopenta[c]c[c] obtained in the previous step. Pyrrole-2-carboxamide 6b was dispersed in 10 mL of diethyl ether, and 2 mL of 0.5 N hydrochloric acid diethyl ether solution was added thereto under ice-cooling, and the precipitate was centrifuged to obtain the title product cis-5-[2-(2-cyanopyrrole). Alkyl-1-yl)-2-oxo-ethylamino]-N-isopropyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide hydrochloride 6 (80 mg, white powder ).

_{^{1 H NMR (CD 3 OD,}} 400MHz) δ 4.70 (m, 1H), 3.92 (m, 2H), 3.76-3.32 (m, 8H), 2.63-1.41 (m, 10H), 1.01 (d, 6H, J =6Hz).

Example 7

Cis-1-(2-(2-(morpholine-4-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethenyl)-2-cyano-pyrrolidine hydrochloride salt

first step

Preparation of 2-(morpholine-4-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-one 7a

The perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate salt 1f (574 mg, 2.4 mmol) was dissolved in 20 mL of acetonitrile with stirring, and potassium carbonate (0.397 g, 2.88 mmol) was added under ice water bath. Morpholine-4-methylhydrazine chloride (0.323 mL, 2.64 mmol) was added dropwise, and the mixture was evaporated, evaporated, evaporated, evaporated, evaporated, evaporated The residue was washed with aq. Hydrogen-cyclopenta[c]pyrrole-5-one 7a (0.572 g, colorless oil), yield 77.3%.

MS m/z (ESI): 239 (MH).

Second step

Cis-1-(2-(2-(morpholine-4-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano-pyrrolidine 7b preparation

2-(morpholine-4-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-one 7a (0.64 g, 2.69 mmol) and 1-(2-aminoethenyl)-2-cyanide The pyridine-pyrrolidine hydrochloride 1d (0.764 g, 4.03 mmol) was dissolved in 50 mL of tetrahydrofuran, and 5 g of sodium sulfate and 0.05 mL of acetic acid were added thereto, and the mixture was reacted at room temperature for 0.5 hour, and then sodium triethoxysulfonate (sodium hydride) was added. 1.71 g, 8.07 mmol), after reacting for 3 hours at room temperature, the reaction mixture was concentrated under reduced pressure, then 50 mL of saturated sodium carbonate solution, and ethyl acetate (50 mL×3) The sodium chloride solution and 50 mL of water were washed, and the ethyl acetate layer was dried over anhydrous magnesium sulfate. (morpholine-4-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethynyl)-2-cyano-pyrrolidine 7b which was used directly in the next reaction.

MS m/z (ESI): 376.7 (M + 1).

third step

Cis-1-(2-(2-(morpholine-4-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethenyl)-2-cyano-pyrrolidine hydrochloride Preparation of salt 7

The cis-1-(2-(2-(morpholine-4-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano group obtained in the previous step Pyrrolidine 7b was dispersed in 10 mL of diethyl ether, and 2 mL of a 0.5 N solution of hydrochloric acid in diethyl ether was added to an ice bath, and the precipitate was centrifuged to give the title product cis-1-(2-(2-(morpholin-4-carbonyl)). -hydrogen-cyclopenta[c]pyrrole-5-ylamino)ethenyl)-2-cyano-pyrrolidine hydrochloride 7 (30 mg, white powder), 3% yield.

Example 8

Cis-1-(2-(2-(pyrrolidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethenyl)2-cyano-pyrrolidine hydrochloride

first step

Preparation of 2-(pyrrolidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-one 8a

Dissolve perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate 1f (478 mg, 2 mmol) in 20 mL of dichloromethane with stirring and add pyrrolidine-1-methylhydrazine chloride (0.276 mL). , 2.5 mmol) and triethylamine (0.84 mL, 6 mmol), react at room temperature overnight, add 10% citric acid solution to adjust pH 4, extract with dichloromethane (50 mL × 3), and combine the organic phase. The residue was washed with 50 mL of aq. -carbonyl)-perhydro-cyclopenta[c]pyrrole-5-one 8a (0.26 g, colorless oil), yield 58.5%.

MS m/z (ESI): 223 (MH).

Second step

Cis-1-(2-(2-(pyrrolidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl)-2-cyano-pyrrolidine 8b preparation

2-(Pyrrolidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-one 8a (0.26 g, 1.17 mmol) and 1-(2-aminoethenyl)-2-cyanide Base-pyrrolidine hydrochloride 1d (0.33 g, 1.75 mmol) was dissolved in 50 mL of tetrahydrofuran, and 5 g of sodium sulfate and 0.05 mL of acetic acid were added thereto. After reacting for 0.5 hour at room temperature, sodium triethoxysulfonate hydride ( 0.75 g, 3.5 mmol), after reacting for 3 hours at room temperature, the reaction mixture was concentrated under reduced pressure, and then 50 mL of saturated sodium carbonate solution was taken and extracted with ethyl acetate (50 mL×3). The sodium chloride solution and 50 mL of water were washed, and the ethyl acetate layer was dried over anhydrous magnesium sulfate. (Pyrrolidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethynyl)2-cyano-pyrrolidine 8b which was used directly in the next reaction.

MS m/z (ESI): 396 (MH).

third step

Cis-1-(2-(2-(pyrrolidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethenyl)2-cyano-pyrrolidine hydrochloride Preparation of 8

The cis-1-(2-(2-(pyrrolidin-1-carbonyl)-perhydro-cyclopenta[c]pyrrole-5-ylamino)ethyl) 2-cyano group obtained in the previous step- The pyrrolidine 8b was dispersed in 10 mL of diethyl ether, and 2 mL of a 0.5 N solution of hydrochloric acid in diethyl ether was added thereto under ice-cooling, and the precipitate was centrifuged to obtain the title product cis-1-(2-(2-(pyrrolidin-1-carbonyl)- All hydrogen-cyclopenta[c]pyrrole-5-ylamino)ethenyl)2-cyano-pyrrolidine hydrochloride 8 (90 mg, white powder).

¹ H NMR (CD ₃ OD, 400 MHz) δ 4.72 (m, 1H), 4.09 (m, 2H), 3.43-3.30 (m, 11H), 2.62 (m, 2H), 2.35 (m, 2H), 2.18 ( m, 2H), 2.08 (m, 2H), 1.77 (m, 4H).

Example 9

Cis-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2 -carbamidine trifluoroacetate

The cis-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethoxyethyl)-N,N-dimethyl-perhydro-cyclopentayl group obtained in Example 1 was obtained. [c] Pyrrole-2-carboxamide 1h (157 mg, 0.54 mmol) was dispersed in 10 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added and stirred for 0.5 hour in an ice bath, and the precipitate was centrifuged to give the title product. -5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethoxyethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2- Formamide trifluoroacetate 9 (201 mg, white powder).

¹ H NMR (CDCl ₃ , 400 MHz) δ 4.74 (t, 1H, J = 5.2 Hz), 3.98 (d, 1H, J = 15.6 Hz), 3.79 (d, 1H, J = 15.6 Hz), 3.57 - 3.25 ( m, 7H), 2.75 (s, 6H), 2.55 (m, 2H), 2.33 (m, 2H), 2.20-2.08 (m, 4H), 1.74 (m, 2H).

Example 10

Trans-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2 -carbamidine trifluoroacetate

first step

Preparation of cis-5-hydroxy-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 10a

N,N-Dimethyl-5-oxo-perhydro-cyclopenta[c]pyrrole-2-carboxamide 1 g (1.58 g, 8.06) in a dry three-necked flask under nitrogen. Ment) dissolved in 30 mL of tetrahydrofuran with stirring, cooled to -25 ° C, and at this temperature, 30 mL of a solution of hydrogenated lithium trit-butoxide aluminum (2.45 g, 9.6 mmol) in tetrahydrofuran was added dropwise at -25 After reacting for 2.5 hours at °C, add water to stop the reaction, add 20 mL of saturated ammonium chloride solution, naturally raise to room temperature, separate the liquid, extract the aqueous phase with dichloromethane (50 mL×3), combine the organic phase, use 50 mL The organic layer was washed with a saturated aqueous solution of sodium chloride, and filtered, filtered, filtered, and evaporated. All hydrogen-cyclopenta[c]pyrrole-2-carboxamide 10a (1.27 g, colorless oil), yield 80%.

MS (m/z) (ESI): 195 (MH).

Second step

Preparation of [cis-2-dimethylaminecarbamyl-perhydro-cyclopenta[c]pyrrole-5-yl]ester 10b

cis-5-Hydroxy-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 10a (1.69 g, 8.5 mmol) in a dry single-necked flask under nitrogen. Dissolved in 30 mL of dichloromethane with stirring, cooled to -5 ° C to 0 ° C with an ice salt bath, and then added triethylamine (1.66 g, 14.45 mmol) and methyl sulfonium chloride (2.2 g, 21.74). After stirring for 0.5 hours, it was naturally warmed to room temperature, and the mixture was reacted at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure. EtOAc (EtOAc m. The title compound was methanesulfonic acid [cis-2-dimethyl]. The residue was purified by chromatography. Aminomethylmercapto-perhydro-cyclopenta[c]pyrrole-5-yl]ester 10b (1.94 g, white solid), yield 83%.

MS (m/z) (ESI): 277 (M+1)

third step

Trans-5-(1,3-di- oxy-1,3-dihydro-isoindol-2-yl)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-A Preparation of guanamine 10c

Under the protection of nitrogen, methanesulfonic acid [cis-2-dimethylamine-mercapto-perhydro-cyclopenta[c]pyrrole-5-yl]ester 10b (1 g, in a dry single-mouth bottle 3.6 mmol) was dissolved in 20 mL of N,N-dimethylformamide with stirring, and potassium phthalate (993 mg, 5.4 mmol) was added, and the temperature was raised to 70 ° C for 3 hours. The N,N-dimethylformamide solution was concentrated, added with 20 mL of water, and extracted with ethyl acetate (50 mL×3). The organic phase was combined, washed with 50 mL of saturated sodium chloride and dried over magnesium sulfate. Filtration and concentration of the filtrate under reduced pressure afforded the title product, <RTI ID=0.0> Hydrogen-cyclopenta[c]pyrrole-carboxamide 10c (1.06 g, white solid), yield 90%.

MS (m/z) (ESI): 328 (M + 1).

the fourth step

Preparation of trans-5-amino-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 10d

In the single-mouth bottle, trans-5-(1,3-di-oxy-1,3-dihydro-isoindol-2-yl)-N,N-dimethyl-perhydro-cyclopenta[ c] Pyrrole-carbamide 10c (1 g, 3.06 mmol) was dissolved in 20 mL of 95% ethanol with stirring, and hydrazine (490 mg, 15.3 mmol) was added, and the mixture was heated to reflux for 8 hours, cooled to room temperature, and filtered. The filtrate was concentrated under reduced pressure to dryness crystals crystals crystals crystalssssssssssssssssssssssssssssssssssssss , N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 10d (290 mg, colorless oil), yield 48%.

MS (m/z) (ESI): 198 (M+1).

the fifth step

Trans-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2 -Preparation of methotrexate 10e

Add de-1-(2-chloro-ethenyl)-2-cyano-pyrrolidine (334 mg, 1.94 mmol) to a dry one-neck flask and add 20 mL of trans-5-amino-N,N- a solution of dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 10d (290 mg, 1.46 mmol) in dichloromethane, heated under reflux for 48 hr. The obtained residue was purified by an analytical method to give trans-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoxyethylamine)-N,N-dimethyl-perhydro - cyclopenta[c]pyrrole-2-carboxamide 10e , which was used directly in the next reaction.

Step 6

Trans-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2 -Preparation of methotrexate trifluoroacetate 10

The trans-5-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethylamino)-N,N-dimethyl-perhydro-cyclopenta[B] obtained in the previous step. c] Pyrrole-2-carboxamide 10e was dissolved in 10 mL of dichloromethane with stirring, 2 mL of trifluoroacetic acid was added and stirred for 0.5 hour to obtain trans-5-(2-(2-cyanopyrrolidine-1-) 2-(Ethyloxyethylamino)-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide trifluoroacetate 10 (201 mg, white solid).

MS (m/z) (ESI): 356 (M + 1).

_{^{1 H NMR (CDCl 3, 400MHz}} ) δ 4.65 (m, 1H), 3.93 (d, 1H, J = 15.2Hz), 3.74 (d, 1H, J = 15.2Hz), 3.69-3.19 (m, 7H), 2.77 (s, 6H), 2.18-1.96 (m, 10H).

Example 11

5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole 2-carbamamine p-toluenesulfonate

first step

Preparation of 5-methylene-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

Potassium tert-butoxide (7.17 g, 0.064 mol) and methyltriphenylphosphonium iodide (25.8 g, 0.064 mol) were dissolved in 150 mL of toluene with stirring under nitrogen, and heated to reflux for 3 hours. At room temperature, add 30 mL of N,N-dimethyl-5-oxo-perhydro-cyclopenta[c]pyrrole-2-carboxamide 1g (5.0 g, 0.0255 mol) in toluene for 30 minutes. Thereafter, the reaction was followed by thin layer chromatography, and the starting material was evaporated. <RTI ID=0.0>>> After suction filtration, the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to give the title product 5-methylene-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 11a (4.0 g, light yellow oily clear liquid), yield: 80%.

MS m/z (ESI): 195.2 (M + 1).

Second step

Preparation of 5-isocyano-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Methylene-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 11a (1.6 g, 8.23 mmol) was dissolved in 30 under a nitrogen atmosphere. Trichloromethane cyanide (4.08 g, 41.2 mmol) and silver perchlorate (5.12 g, 24.7 mmol) were added to dichloromethane, and stirred at room temperature overnight, then partially reacted with saturated sodium hydrogen carbonate solution. The liquid was traced by thin layer chromatography. Most of the raw materials remained. Under ice bath, 20 mL of saturated sodium bicarbonate solution was added, and the reaction was exothermic. After 10 minutes, it was filtered to remove solids. After liquid separation, use acetic acid. The ester was extracted (50 mL×4), EtOAcjjjjjjjjjjjjjjjjjj Trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 11b (0.33 g, pale yellow oil), yield: 18.1%.

GC-MS: 221.1 (M ⁺ ).

_{^{1 H NMR (CDCl 3, 400MHz}} ) δ 3.31 (m, 4H), 2.97 (m, 2H), 2.84 (s, 6H), 2.32 (m, 2H), 1.52 (s, 3H), 1.46 (m, 2H ).

third step

Preparation of 5-amino-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

0.38 mL of 6N hydrochloric acid was added to 5-isocyano-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 11b (0.388 g, under ice bath). 1.75 mmol) in 15 mL ethanol solution, remove the ice bath, react at room temperature for 1.5 hours, trace the reaction by thin layer chromatography, disappear the raw materials, add 20 mL of saturated sodium bicarbonate solution to stop the reaction, use dichloro The methane (50 mL×3) was extracted, the organic phase was combined, and the solvent was evaporated to dryness and purified to the title product product 5-amino-N,N,5-trimethyl-perhydro-cyclopenta [c] Pyrrole-2-carboxamide 11c (0.28 g, yellow oil), yield: 75.7%.

MS m/z (ESI):21.21.

the fourth step

5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole Preparation of 2-carbamamine

5-Amino-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 11c (200 mg, 0.92 mmol) was dissolved in 8 mL of N with stirring. To N-dimethylformamide, 1-(2-chloro-ethenyl)-2-cyano-pyrrolidine (175 mg, 1.02 mmol) was added, and the reaction was carried out at room temperature overnight at 40-50 ° C. In a water bath, N,N-dimethylformamide was removed under reduced pressure and purified by silica gel column chromatography to give the title product 5-[2-(2-cyano-pyrrolidin-1-yl)-2- Side oxy-ethylamino]-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 11d (135 mg, colorless oily liquid), yield: 42.3 %. 5-Amino-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 11c (130 mg) was recovered.

MS m/z (ESI): 348.2 (M + 1).

the fifth step

5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole Preparation of 2-carbamamine p-toluenesulfonate

5-[2-(2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro-ring under ice bath Ethylene [c]pyrrole-2-carboxamide 11d (20 mg, 0.057 mmol) and p-toluenesulfonic acid monohydrate (12 mg, 0.063 mmol) were dissolved in 2 mL of dichloromethane with stirring and stirred for 10 min. After that, the ice bath was removed, and the solvent was evaporated to dryness crystals crystals crystals crystals crystals crystals 2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide p-Toluenesulfonate 11 (26 mg, white solid), yield: 86.8%.

MS m/z (ESI): 348.2 (M + 1).

_{^{1 H NMR (CDCl 3, 400MHz}} ) δ 7.68 (d, 2H), 7.17 (d, 2H), 3.72 (m, 1H), 3.68 (m, 1H), 3.58 (m, 1H), 3.30 (m, 4H ), 2.98 (s, 2H), 2.81 (s, 6H), 2.55 (m, 2H), 2.22 (m, 4H), 2.05 (m, 2H), 1.62 (m, 2H), 1.56 (s, 3H) .

Example 12

5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole 2-carbamamine p-toluenesulfonate

first step

Preparation of 5-cyano-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

N,N-Dimethyl-5-oxo-perhydro-cyclopenta[c]pyrrole-2-carboxamide 1 g (12.9 g, 0.066 mol) and p-toluenesulfonate under ice bath The isomeric nitrile (14.2 g, 0.0727 mol) was dissolved in 240 mL of ethylene glycol dimethyl ether with stirring, and a solution of potassium terp-butoxide (14.8 g, 0.132 mol) in a third butanol was added dropwise, and the mixture was removed. The mixture was stirred at room temperature overnight, and traced by thin layer chromatography. When the reaction was completed, 100 mL of water, 50 mL of saturated sodium chloride solution was added, and extracted with ethyl acetate (200 mL×3), and the organic phase was combined. Drying over anhydrous sodium sulfate, suction filtration, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to give the title product 5-cyano-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2 -Procarbamide 12a (7.0 g, pale yellow oily), yield: 51%.

MS m/z (ESI): 208.1 (M + 1).

¹ H NMR (DMSO-D ₆ , 400 MHz) δ 3.5-3.0 (m, 4H), 2.75 (s, 6H), 2.6 (m, 1H), 2.1-1.5 (m, 6H).

Second step

Preparation of 5-cyano-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Cyano-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12a (4.2 g, 20.2 mmol) and methyl iodide (11.5 g, under N2) 80.8 mmol) was dissolved in 100 mL of anhydrous tetrahydrofuran under stirring. At room temperature, lithium hexamethyldidecylamine (80.8 mL, 80.8 mmol) was added dropwise, and the reaction was carried out for 2 hours at room temperature. 100 mL of water, extracted with ethyl acetate (200 mL × 3), dried over anhydrous sodium sulfate, filtered, filtered, and evaporated. Purified by column chromatography, the following point is 5-cyano-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12b (2.411 g, pale yellow oil Shape).

MS m/z (ESI): 2221.

_{^{1 H NMR (CDCl 3, 400MHz}} ) δ 3.44 (m, 2H), 3.3 (m, 2H), 2.85 (s, 6H), 2.77 (m, 2H), 2.06 (m, 2H), 2.00 (m, 2H ), 1.36 (s, 3H).

third step

Preparation of 2-dimethylamine-mercapto-5-methyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid

5-Cyano-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12b (6.99 g, 31.6 mmol) was dissolved in 90 mL of 36% with stirring In hydrochloric acid, the mixture was stirred at 50 ° C for 48 hours in an oil bath, and the reaction was followed by thin layer chromatography. When the starting material disappeared, 150 mL of water was added at the end of the reaction, and the reaction solution was neutralized with potassium carbonate solid in an ice bath to adjust the pH. The mixture was extracted with ethyl acetate (150 mL×3), and extracted with dichloromethane (150 mL×3). 2-Dimethylamine-mercapto-5-methyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid 12c (7.0 g, yellow liquid), yield: 92%, which was used directly under One step reaction.

MS m/z (ESI): 242.

the fourth step

Preparation of 5-amino-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

2-Dimethylamine-mercapto-5-methyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid 12c (1.0 g, 4.2 mmol) was dissolved in 25 under ice-cooling. Triethylamine (463.5 mg, 4.58 mmol) and ethyl chloroformate (497 mg, 4.58 mmol) in 10 mL of acetone were added dropwise at -5 ° C in acetone for 15 minutes at -5 °C. A 10 mL aqueous solution of sodium azide (546 mg, 8.4 mmol) was added, and the reaction was continued at -5 °C for 30 minutes. The reaction was quenched by the addition of 25 mL of water and extracted with ethyl acetate (50 mL×3). Drying, suction filtration, and concentration of the filtrate under reduced pressure afforded crude product-5-(azidocarbonyl)-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12d (700 mg, yellow oil).

Fifth to sixth steps

Add the yellow oil 5-(azidocarbonyl)-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12d (700 mg) to 20 mL The mixture was refluxed for 2 hours in toluene, and toluene was evaporated to give the product (3aS, 5r, 6aR)-5-isocyano-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2 ( 1H)-carbalamine 12e , 8 mL of 8N hydrochloric acid was added dropwise under ice bath, stirred for 30 minutes, traced by thin layer chromatography until the reaction material disappeared, and adjusted to pH>12 with 3N sodium hydroxide in an ice bath. The mixture was extracted with ethyl acetate (15 mL×3). EtOAcjjjjjjjjjj , 5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12f (500 mg, pale yellow oily liquid).

MS m/z (ESI):21.21.

¹ H NMR (DMSO-D ₆ , 400 MHz) δ 3.5-3.0 (m, 4H), 2.72 (s, 6H), 1.7-1.3 (m, 6H), 1.04 (s, 3H).

Seventh step

5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole Preparation of 2-carbamamine

5-Amino-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12f (332 mg, 1.57 mmol) was dissolved in 10 mL of N with stirring. To a mixed solvent of N-dimethylformamide/CH ₂ Cl ₂ (1:1), 1-(2-chloro-ethenyl)-2-cyano-pyrrolidine (217 mg, 1.26 mmol) was added. After reacting at room temperature overnight, the solvent was evaporated and purified with EtOAc EtOAc EtOAc. Base]-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12 g (240 mg, colorless oil), yield: 55%.

MS m/z (ESI): 348.2 (M + 1).

Eighth step

5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl Preparation of thio-perhydro-cyclopenta[c]pyrrole-2-carboxamide p-toluenesulfonate

5-[2-(2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro-cyclopenta[c] Pyrrole-2-carboxamide 12g (150 mg, 0.43 mmol) and p-toluenesulfonic acid monohydrate (82 mg, 0.43 mmol) were dissolved in 4 mL of dichloromethane with stirring, stirred for 10 min and evaporated to dryness Solvent in the solvent gave the title product 5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro- Cyclopenta[c]pyrrole-2-carboxamide p-toluenesulfonate 12 (220 mg, pale yellow solid), yield: 95.3%.

MS m/z (ESI): 348.2 (M + 1).

_{^{1 H NMR (DMSO-D 6}} , 400MHz) δ 4.82 (m, 1H), 3.97 (s, 2H), 3.79 (m, 2H), 3.49 (m, 2H), 3.21 (m, 4H), 2.75 (s , 6H), 2.62 (m, 2H), 2.19 (m, 2H), 1.92-1.61 (m, 3H), 2.06 (m, 3H), 1.2 (s, 3H).

Example 13

5-(2-(2-cyano-4-fluoropyrrolidin-1-yl)-2-oxoethylamino)-N,N,5-trimethyl-perhydro-cyclopenta [c]pyrrole-2-carboxamide tartrate

first step

Preparation of (2S,4R)-1-(t-butoxycarbonyl)-4-hydroxypyrrolidine-2-carboxylic acid

Hydroxyproline 13a (60 g, 0.458 mol) was added to a 750 mL tetrahydrofuran/water (2:1) mixed solvent, 252 mL of 10% sodium hydroxide solution was added, and 750 mL of di-tert-butyl dicarbonate was added. (136 g, 0.624 mol) in tetrahydrofuran/water (2:1) solution, react at room temperature overnight, add 500 mL of ethyl acetate, separate the layers, discard the organic layer, and adjust the pH to 2 with concentrated hydrochloric acid. 1.5 L of ethyl acetate was extracted, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate. Pyrrolidine-2-carboxylic acid 13b (86.4 g, colorless oil), yield: 80%.

Second step

Preparation of (2S,4R)-2-aminoformamido-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester

(2S,4R)-1-(Tertixobutoxycarbonyl)-4-hydroxypyrrolidine-2-carboxylic acid 13b (86.4 g, 0.374 mol) was dissolved in 1.2 L of tetrahydrofuran under stirring in an argon atmosphere. Triethylamine (41 g, 0.411 mol), cooled to -15 ° C, added ethyl chloroformate (43.84 g, 0.411 mol), stirred for 10 minutes, then added 236.8 mL of ammonia water, slowly warmed to 5 ° C within 2 hours Then add 32 g of ammonium chloride, stir for 30 minutes, stop, separate the liquid, the organic phase is dried over anhydrous sulfuric acid, the aqueous phase is extracted with ethyl acetate (100 mL×2), the organic phase is combined, the solvent is evaporated, and dried in vacuo. The title compound (2S,4R)-2-amine-carbazyl-4-hydroxypyrrolidine-l-carboxylic acid tert-butyl ester 13c (74 g, white solid).

third step

Preparation of (2S,4R)-2-cyano-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester

(2S,4R)-2-Aminoformamido-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester 13c (74 g, 0.3217 mol) was dissolved in 740 mL of pyridine under stirring, and cooled. To -20 ° C, trifluoroacetic anhydride (169 g, 0.804 mol) was added dropwise, and the mixture was added dropwise to room temperature. The reaction was traced by thin layer chromatography until the starting material disappeared. Water was added to stop the reaction, and 0.8 L of ethyl acetate was added. Stir, separate the liquid, wash the organic phase with 500 mL of saturated sodium chloride solution, neutralize the pyridine with 400 mL of concentrated hydrochloric acid to slightly acidic, wash with 300 mL of 2M sodium hydroxide solution, and wash with 500 mL of saturated sodium chloride solution. , dried over anhydrous magnesium sulfate, suction filtered, the filtrate was concentrated under reduced pressure to give the title product (2S, 4R) -2- cyano-4-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester 13d (49.7g, brown oil (form), yield: 73%.

the fourth step

Preparation of (2S,4S)-2-cyano-4-fluoropyrrolidine-1-carboxylic acid tert-butyl ester

(2S,4R)-2-Cyano-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester 13d (49.7 g, 0.2344 mol) was dissolved in 1130 mL of dichloroethane under stirring in an argon atmosphere. After cooling to -30 ° C, diethylphosphonium trifluoride (56.7 g, 0.3516 mol) was added, and after reacting for 45 minutes, it was raised to -5 ° C, naturally raised to room temperature, and reacted overnight, and the reaction was followed by thin layer chromatography. When the raw material disappears, slowly add saturated sodium bicarbonate solution, vigorously bubbling, cooling, and keep the temperature below 20 °C until the pH of the reaction solution is greater than 7, add ice water, add 500 mL of dichloromethane, separate the liquid, and then use 500 The title product (2S, 4S)-2-cyanide is obtained by washing with saturated sodium hydrogen sulfate solution and 500 mL of saturated sodium chloride solution, dried over anhydrous magnesium sulfate, suction filtration, and the filtrate is concentrated under reduced pressure. Base 4-fluoropyrrolidine-1-carboxylic acid tert-butyl ester 13e (50 g, brownish yellow solid), yield: 100%.

the fifth step

Preparation of (2S,4S)-4-fluoro-2-cyanopyrrolidine

(2S,4S)-2-Cyano-4-fluoropyrrolidine-1-carboxylic acid tert-butyl ester 13e (1 g, 4.6 mmol) was dissolved in 2 mL of ethyl acetate under stirring. Cool to 15 ° C, add 2.5 mL of 3M hydrogen chloride 1,4-two The alkane solution was stirred at room temperature for 2 hours, and reacted at 25 ° C or lower for 5 hours. The thin layer chromatography showed that the resulting reaction solution contained a raw material, and the turbid reaction solution was filtered, and the mother liquid was further stirred at room temperature for 2 hours, and white again. The solid was precipitated, filtered, and the m~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ : 76.6%.

Step 6

Preparation of (2S,4S)-1-(2-chloroethyl)-4-fluoro-2-cyanopyrrolidine

Chloroacetate chloride (11.13 g, 98.5 mmol) was dissolved in 120 mL of dichloromethane under stirring, cooled to 0 ° C, (2S,4S)-4-fluoro-2-cyanopyrrole Alkane 13f (11.4 g, 75.7 mmol) was dissolved in 400 mL of dichloromethane with stirring, triethylamine (16.1 g, 158.97 mmol) was added, and the solution was added dropwise to the chloroethyl chloride in 30 minutes. In a solution of methyl chloride, react at 0 ° C for 2 hours, add 200 mL of water, 150 mL of dichloromethane, extract and separate the organic phase with a small amount of saturated sodium bisulfate solution to neutral, then use 300 mL of water and 300 mL. The residue was washed with a saturated aqueous solution of sodium chloride and dried over anhydrous sodium sulfate -4-fluoro-2-cyanopyrrolidine 13 g (8 g, white crystal), yield: 60%.

Reference: WO2003002553.

Seventh step

5-(2-(2-cyano-4-fluoropyrrolidin-1-yl)-2-oxoethylamino)-N,N,5-trimethyl-perhydro-cyclopenta[ c] Preparation of pyrrole-2-carboxamide

5-Amino-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12f (130 mg, 0.616 mmol), (2S, 4S) 1-(2-chloroethenyl)-4-fluoro-2-cyanopyrrolidine 13g (117.4 mg, 0.616 mmol) and potassium carbonate (85 mg, 0.616 mmol) were dissolved in 2 mL dichloromethane with stirring The reaction was carried out in a mixed solvent of 2 mL of N,N-dimethylformamide at room temperature overnight, and the reaction was followed by thin layer chromatography, the starting material was substantially disappeared, and the dichloromethane and N in the reaction mixture were removed under reduced pressure. , N-dimethylformamide, the residue obtained was purified by silica gel column chromatography to give the title product 5-(2-(2-cyano-4-fluoropyrrolidin-1-yl)-2-one Oxyethylamino)-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2(1H)-carbenamide 13h (0.13 g, colorless oily liquid), yield : 58%.

Eighth step

5-(2-(2-Cyano-4-fluoropyrrolidin-1-yl)-2-oxoethylamino)-N,N,5- Preparation of trimethyl-perhydro-cyclopenta[c]pyrrole-2(1H)-carbenamide tartrate

5-(2-(2-Cyano-4-fluoropyrrolidin-1-yl)-2-oxoethylamino)-N,N,5-trimethyl-perhydrogen at room temperature -cyclopenta[c]pyrrole-2(1H)-carbenamide 13 (0.16 g, 0.44 mmol) was dissolved in 5 mL of dichloromethane with stirring, and 5 mL of tartaric acid (65.6 mg, 0.44 mmol) was added dropwise. The acetone solution was reacted at room temperature for 30 minutes, and a white solid was precipitated, which was filtered, and the solid was washed with acetone to give the title product 5-(2-(2-cyano-4-fluoropyrrolidin-1-yl)-2-one Oxyethylamino)-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide tartrate 13 (0.18 g, white solid), yield: 82% .

_{^{1 H NMR (DMSO-D 6}} , 400MHz) δ 5.76 (m, 1H), 5.46 (m, 1H), 5.0 (m, 1H), 4.08-4.05 (m, 4H), 3.97 (m, 2H), 3.69 (m, 2H), 2.73 (s, 6H), 2.61 (m, 2H), 1.87 (m, 3H), 1.57 (m, 3H), 1.18 (s, 3H), 1.9 (m, 2H).

Example 14

5-benzyl-5-[2-(2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro -cyclopenta[c]pyrrole-2-carboxamide tartrate

first step

Preparation of 5-benzyl-5-cyano-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Cyano-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12a (4.0 g, 19.3 mmol) was dissolved in 60 mL of tetrahydrofuran under stirring, and benzyl chloride (5.4 g) was added. , 42.5 mmol), lithium hexamethyldidecylamine (42.5 mL, 42.5 mmol) was added dropwise with stirring at room temperature, and stirred at room temperature for 4 hours. The reaction was traced by thin layer chromatography until the starting material disappeared, and 50 mL was added. The reaction was quenched with water, EtOAc (EtOAc m.jjHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH -Cyano-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 14a (2.6 g, pale yellow solid), yield: 46%.

Second step

Preparation of 5-benzyl-5-methylindenyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Benzyl-5-cyano-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 14a (2.0 g, 6.7 mmol) was dissolved in 100 mL with stirring Di-isobutylaluminum (20.2 mL, 20.2 mmol) was hydrogenated at 0 ° C in dichloromethane, and the reaction was followed by thin layer chromatography until the disappearance of the starting material. The reaction was stopped with water and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, filtered, and evaporated. Hydrogen-cyclopenta[c]pyrrole-2-carboxamide 14b (729 mg, colorless oily liquid), yield: 36%.

third step

Preparation of 5-benzyl-2-dimethylaminomethylindenyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid

Under a nitrogen atmosphere, 5-benzyl-5-methylindenyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 14b (0.729 g, 2.43 mmol) was stirred. Dissolved in a mixed solvent of 28 mL of tetrahydrofuran and 14 mL of water, and added sodium dihydrogen phosphate dihydrate (1.14 g, 7.29 mmol), sodium chlorite (0.66 g, 7.29 mmol) and 2-A at 0 °C. Base-2-butene (0.513 g, 7.32 mmol), reacted at 0 ° C for 2 hours, the reaction was traced by thin layer chromatography until the disappearance of the starting material, and the mixture was extracted with ethyl acetate. The organic phase was combined and dried over anhydrous magnesium sulfate. After suction filtration, the filtrate was concentrated under reduced pressure, and the obtained residue was purified to silica gel column chromatography to give the title product 5-benzyl-2-dimethylaminocarbazyl-perhydro-cyclopenta[c]pyrrole. -5-carboxylic acid 14c (0.76 g, colorless oily liquid), yield: 98%.

MS m/z (ESI): 317.3 (M + 1).

_{^{1 H NMR (DMSO-D 6}} , 400MHz) δ 7.5-7.0 (m, 5H), 3.24 (m, 2H), 3.1 (m, 2H), 2.76 (s, 2H), 2.7 (s, 6H), 2.68 (m, 2H), 1.99-1.55 (m, 4H).

the fourth step

Preparation of 5-benzyl-2-dimethylaminomethylmercapto-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide

5-benzyl-2-dimethylaminocarbazyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid 14c (0.86 g, 2.72 mmol) was dissolved in 30 mL with stirring under ice-cooling In acetone, triethylamine (0.303 g, 2.99 mmol), 15 mL of ethyl chloroformate (0.325 g, 2.99 mmol) in acetone were added dropwise at -5 ° C, stirred for 15 minutes, and 15 mL of azide was added dropwise. Aqueous solution of sodium (0.353 g, 5.44 mmol) was stirred for 30 minutes. The reaction was evaporated to dryness eluting with EtOAc (EtOAc). The filtrate was concentrated under reduced pressure to give the title product: 5-benzyl-2-dimethylaminocarbazyl-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide 14d (0.9 g, pale yellow oil Liquid), yield: 97%.

the fifth step

Preparation of 5-benzyl-5-isocyanato-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Benzyl-2-dimethylaminocarbazyl-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide 14d (1.0 g, 2.72 mmol) was dissolved in 20 mL of toluene with stirring The mixture was heated under reflux for 1.5 hours, and toluene was concentrated. The residue obtained was obtained from the title product 5-benzyl-5-isocyanyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 14e . In the next step.

Step 6

Preparation of 5-amino-5-benzyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

To the 5-benzyl-5-isocyanato-perhydro-cyclopenta[c]pyrrole-2-carboxamide 14e obtained in the previous step, 12 mL of 8 N hydrochloric acid was added dropwise and stirred at room temperature. After 30 minutes, the pH was adjusted to 9 with 4 N sodium hydroxide solution, extracted with ethyl acetate, and the organic phase was combined, dried over anhydrous sodium sulfate, filtered, filtered, and the filtrate was concentrated under reduced pressure. To give the title product 5-amino-5-benzyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 14f (0.55 g, colorless oily). Yield: 70%.

Seventh step

5-benzyl-5-[2-(2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro -Preparation of cyclopenta[c]pyrrole-2-carboxamide

5-Amino-5-benzyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 14f (0.1 g, 0.35 mmol) was dissolved in 4 mL with stirring (2S,4S)-1-(2-chloroethenyl)-4-fluoro-2 was added to a mixed solvent of dichloromethane/N,N-dimethylformamide (V/V=1/1). -Cyanopyrrolidine 13g (66.5 mg, 0.35 mmol) and potassium carbonate (49 mg, 0.35 mmol), reacted at 40 ° C for 12 hours, traced the reaction by thin layer chromatography until the starting material disappeared, concentrated the reaction mixture, using a rubber tube column The obtained residue was purified by chromatography to give the titled product 5-benzyl-5-[2-(2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino] -N,N-Dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 14 g (87 mg, white solid), yield: 56%.

MS m/z (ESI): 4421.

Eighth step

5-benzyl-5-[2-(2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro -cyclopenta[c]pyrrole-2-carboxamide tartrate Preparation

5-Benzyl-5-[2-(2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-all Hydrogen-cyclopenta[c]pyrrole-2-carboxamide 14g (87 mg, 0.197 mmol) was dissolved in 3 mL of dichloromethane with stirring, 3 mL of tartaric acid in acetone was added dropwise, and reacted for 30 minutes at room temperature. The solvent was concentrated, and ethyl acetate and acetone were added to recrystallize to give the title product 5-benzyl-5-[2-(2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxoxy Benzyl-ethylamino]-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide tartrate 14 (80 mg, white solid), yield: 68%.

_{^{1 H NMR (DMSO-D 6}} , 400MHz) δ 7.32-7.2 (m, 5H), 5.55 (d, 1H), 5.41 (d, 1H), 4.97 (m, 1H), 4.31 (s, 2H), 4.08 (m, 1H), 4.0-3.5 (m, 6H), 2.81 (s, 2H), 2.78 (s, 6H), 2.5 (m, 2H), 2.0 (m, 3H), 1.6-1.3 (m, 6H) ).

Example 15

5-cyclohexylmethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-ring Pentyl[c]pyrrole-2-carboxamide tartrate

first step

Preparation of 5-cyano-5-cyclohexylmethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Cyano-perhydro-N,N-dimethyl-cyclopenta[c]pyrrole-2-carboxamide 12a (3.66 g, 17.6 mmol) was dissolved in 150 mL of tetrahydrofuran with stirring and dripped Bromomethylcyclohexane (6.2 g, 35.2 mmol), lithium hexamethyldidecylamine (35.2 mL, 35.2 mmol) was added dropwise, and reacted at room temperature for 2 hours. The reaction was traced to the starting material by thin layer chromatography. After disappearing, 150 mL of saturated ammonium chloride solution was added, and the mixture was extracted with ethyl acetate (60 mL×3). The organic phase was combined, dried over anhydrous magnesium sulfate, filtered, filtered, and the filtrate was concentrated under reduced pressure. To give the title product 5-cyano-5-cyclohexylmethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 15a (2.2 g, yellow solid). Yield: 41.5%.

MS m/z (ESI): 304.5 (M + 1).

Second step

Preparation of 5-cyclohexylmethyl-5-methylindenyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Cyano-5-cyclohexylmethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 15a (1.2 g, 3.95 mmol) at 0 °C Dissolve in 30 mL of dichloromethane under stirring, dilute hydrogenated diisobutylaluminum (11.8 mL, 11.8 mmol) in an ice bath, react for 1 hour, trace the reaction by thin layer chromatography until the starting material disappears, add 100 mL of saturation. Potassium sodium tartrate solution, stirred until the solution was transparent, extracted with ethyl acetate (100 mL×3), and the organic phase was combined, dried over anhydrous magnesium sulfate, filtered, filtered, and the filtrate was concentrated under reduced pressure and purified by gel column chromatography. To give the title product 5-cyclohexylmethyl-5-carbamimidyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 15b (0.4 g, slightly yellow oil ()), yield: 33%.

MS m/z (ESI): 307.4 (M + 1).

third step

Preparation of 5-cyclohexylmethyl-2-dimethylaminomethylindenyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid

Dissolving 5-cyclohexylmethyl-5-methylindenyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 15b (0.713 g, 2.33 mmol) with stirring To a mixed solvent of 60 mL of tetrahydrofuran and 30 mL of water, sodium dihydrogen phosphate dihydrate (1.09 g, 6.99 mmol), sodium chlorite (0.79 g, 6.99 mmol) and 2-methyl-2 were added at 0 °C. -butene (0.62 mL, 7.0 mmol), reacted at 0 ° C for 2 hours, followed by thin layer chromatography to trace the disappearance of the starting material, evaporated to dryness of tetrahydrofuran, ethyl acetate was added to extract aqueous phase, and the organic phase was combined with anhydrous magnesium sulfate. The mixture was dried, suction filtered, and the filtrate was evaporated,jjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj c] pyrrole-5-carboxylic acid 15c (0.75 g, yellow solid), yield: 100%.

MS m/z (ESI): 323.3 (M + 1).

the fourth step

Preparation of 5-cyclohexylmethyl-2-dimethylaminomethylmercapto-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide

5-Cyclohexylmethyl-2-dimethylaminomethylindenyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid 15c (0.75 g, 2.33 mmol) was dissolved under stirring in an ice bath Add triethylamine (0.36 mL, 2.56 mmol) at 0 to -5 ° C in 20 mL of acetone, add 2 mL of ethyl chloroformate (0.25 mL, 2.56 mmol) in acetone, stir for 15 minutes, add 2 An aqueous solution of sodium azide (0.303 g, 4.66 mmol) was maintained at a temperature of 0 to -5 ° C, and the reaction was continued for 30 minutes. The reaction was traced by thin layer chromatography until the starting material disappeared, and the acetone in the reaction mixture was evaporated to dryness. The mixture was extracted with EtOAc (EtOAc (EtOAc)EtOAc. Aminomethylmercapto-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide 15d (0.79 g, yellow oil) was used directly in the next step.

the fifth step

Preparation of 5-cyclohexylmethyl-5-isocyanato-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Cyclohexylmethyl-2-dimethylaminomethylindenyl-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide 15d (0.79 g, 2.27 mmol) was dissolved in 20 with stirring The mixture was heated under reflux for 1 hour, and the reaction was traced by thin-layer chromatography to the disappearance of the starting material, and the residue was evaporated to give the titled product: 5-cyclohexylmethyl-5-isocyanyl-N,N- Dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 15e (0.65 g, gray solid) was used directly in the next step.

MS m/z (ESI): 320.4 (M + 1).

Step 6

Preparation of 5-amino-5-cyclohexylmethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

Under ice bath, 10 mL of 8 N hydrochloric acid was added dropwise to 5-cyclohexylmethyl-5-isocyanato-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamidine In the amine 15e (0.65 g, 2.03 mmol), the ice bath was removed, and the reaction was carried out at 50 ° C for 20 minutes. The reaction was traced by thin layer chromatography until the disappearance of the starting material. Concentrated aqueous ammonia was added to the reaction solution to adjust the pH to more than 8. The aqueous layer was extracted with ethyl acetate (30 mL, EtOAc). -5-Cyclohexylmethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 15f (0.5 g, gray oil), yield: 84%.

MS m/z (ESI): 299.

Seventh step

5-cyclohexylmethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-ring Preparation of pentato[c]pyrrole-2-carboxamide

5-Amino-5-cyclohexylmethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 15f (0.115 g, 0.39 mmol) was dissolved in stirring Add 10-methylene chloride/N,N-dimethylformamide (V/V=1/1) to 1-(2-chloro-ethenyl)-2-cyano-pyrrolidine (54 mg , 0.31 mmol), reacted at 50 ° C for 2 hours, the reaction was traced by thin layer chromatography until the disappearance of the starting material, and the dichloromethane and N,N-dimethylformamide in the reaction mixture were removed by concentration under reduced pressure. The obtained residue was purified by column chromatography to give the titled product 5--cyclohexylmethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-yloxy-ethylamino] -N,N-Dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 15 g (113 mg, colorless oily liquid), yield: 67%.

MS m/z (ESI): 430.5 (M + 1).

Eighth step

5-cyclohexylmethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-ring Preparation of pentato[c]pyrrole-2-carboxamide tartrate

5-Cyclohexylmethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro- 15 g (113 mg, 0.263 mmol) of cyclopenta[c]pyrrole-2-carboxamide was dissolved in 10 mL of ethyl acetate with stirring, 2 mL of a solution of tartaric acid in acetone was added dropwise, stirred for 30 minutes, and filtered. The title product 5-cyclohexylmethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro - Cyclopenta[c]pyrrole-2-carboxamide tartrate 15 (40 mg, white solid).

_{^{1 H NMR (DMSO-D 6}} , 400MHz) δ 5.2 (m, 1H), 4.77 (s, 2H), 4.18 (m, 3H), 4.02 (m, 1H), 3.63 (m, 2H), 3.32 (m , 2H), 2.73 (s, 6H), 2.58 (m, 2H), 2.16-1.87 (m, 9H), 1.59-1.19 (m, 8H).

Example 16

5-cyclopentyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopentyl And [c]pyrrole-2-carboxamide tartrate

first step

Preparation of 5-cyano-5-cyclopentyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

Dissolving 5-cyano-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12a in 30 mL of N,N-dimethylformamidine under nitrogen To the amine, iodine cyclopentane (5.4 g, 27.5 mmol) was added, lithium hexamethyldidecylamine (27.5 mL, 27.5 mmol) was added dropwise, and stirred at room temperature for 2 hours, followed by thin layer chromatography. After the reaction was completed, the mixture was evaporated to dryness. EtOAc was evaporated. The resulting residue was purified by column chromatography to give the title product 5-cyano-5-cyclopentyl-yl -N, N- dimethyl - perhydro - cyclopenta [c] pyrrole-2-acyl-amine 16a (2.6 g, pale yellow solid), yield: 42%.

MS m/z (ESI): 266.

¹ H NMR (DMSO-D6, 400 MHz) δ 3.35-3.0 (m, 4H), 2.24 (s, 6H), 1.34-2.5 (m, 15H).

Second step

Preparation of 5-cyclopentyl-5-methylindenyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

Stirring 5-cyano-5-cyclopentyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 16a (0.817 g, 2.97 mmol) at 0 °C Dissolved in 40 mL of dichloromethane, and distilled hydrogenated diisobutylaluminum (8.9 mL, 8.9 mmol) in an ice bath. After reacting for 45 minutes, the reaction was traced by thin layer chromatography until the starting material disappeared. Add 100 mL of saturated sodium potassium tartrate solution, stir until the solution is clear, extract with ethyl acetate (100 mL×4), combine the organic phase, dry over anhydrous magnesium sulfate, suction filtration, and concentrate the filtrate under reduced pressure. The obtained residue was purified by chromatography to give the titled product 5- 5-pentyl-5-carbazyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 16b (0.266 g , light yellow oil), yield: 32%.

MS m/z (ESI): 299.

third step

Preparation of 5-cyclopentyl-2-dimethylaminomethylindenyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid

Dissolving 5-cyclopentyl-5-methylindenyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 16b (0.266 g, 0.955 mmol) in 20 In a mixed solvent of mL tetrahydrofuran and 10 mL of water, sodium dihydrogen phosphate dihydrate (0.448 g, 2.87 mmol), sodium chlorite (0.26 g, 2.87 mmol) and 2-methyl-2- at 0 ° C were added. Butene (0.24 mL, 2.88 mmol) was reacted at 0 °C for 2 hours. The reaction was traced by thin-layer chromatography to dryness of material, evaporated to dryness, and the aqueous phase was extracted with ethyl acetate. The organic phase was combined and dried over anhydrous magnesium sulfate. After suction filtration, the filtrate was concentrated under reduced pressure, and the obtained residue was purified to silica gel column chromatography to give the title product 5-cyclopentyl-2-dimethylaminocarbamoyl-perhydro-cyclopenta[c] Pyrrole-5-carboxylic acid 16c (0.28 g, yellow solid), yield: 99.6%.

MS m/z (ESI): 295.5 (M + 1).

the fourth step

Preparation of 5-cyclopentyl-2-dimethylaminomethylmercapto-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide

5-Cyclopentyl-2-dimethylaminocarbazyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid 16c (0.28 g, 6.95 mmol) was dissolved in 20 under ice-cooling. Triethylamine (0.15 mL, 1.05 mmol), 2 mL of ethyl chloroformate (0.1 mL, 1.05 mmol) in acetone was added dropwise from 0 to -5 ° C, and stirred for 15 min. An aqueous solution of sodium (0.124 g, 1.9 mmol) was further reacted for 30 minutes. The reaction was traced by thin-layer chromatography to dryness of material, evaporated to dryness, and evaporated to ethyl acetate. Drying, suction filtration, and concentrating the filtrate under reduced pressure afforded the title product 5-cyclopentyl-2-dimethylaminocarbazyl-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide 16d ( 0.287 g, yellow oil), yield: 95%, which was used directly in the next step.

the fifth step

Preparation of 5-cyclopentyl-5-isocyanato-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

Dissolving 5-cyclopentyl-2-dimethylaminocarbazyl-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide 16d (0.287 g, 0.9 mmol) in 10 mL of toluene with stirring The mixture was heated under reflux for 1 hour, and the reaction was traced by thin layer chromatography to yield disappearance, and the solvent was evaporated to give the title product 5-cyclopentyl-5-isocyanyl-N,N-dimethyl-perhydro-ring. Pentac[c]pyrrole-2-carboxamide 16e , which was used directly in the next reaction.

MS m/z (ESI): 299.

Step 6

Preparation of 5-amino-5-cyclopentyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

10 parts of 5-cyclopentyl-5-isocyanato-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 16e obtained in the next step under ice bath HCl 8N hydrochloric acid, remove the ice bath, react at 50 ° C for 15 minutes, follow the reaction by thin layer chromatography until the disappearance of the raw materials, add concentrated ammonia water to adjust the pH of the reaction solution to more than 8, extract with ethyl acetate, and combine the organic phase, The organic layer was dried over anhydrous magnesium sulfate, filtered, filtered, evaporated, evaporated,,,,,,,,,,,,, - cyclopenta[c]pyrrole-2-carboxamide 16f (0.18 g, yellow oily liquid), yield: 81.8%.

MS m/z (ESI): 266.2 (M + 1).

Seventh step

5-cyclopentyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamine Preparation of ]-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Amino-5-cyclopentyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 16f (0.108 g, 0.407 mmol) and 1 under nitrogen atmosphere -(2-Chloro-ethenyl)-2-cyano-pyrrolidine (70 mg, 0.407 mmol) was dissolved in 3 mL of N,N-dimethylformamide with stirring, and potassium carbonate (57 mg, 0.407 mmol), the reaction was carried out in an oil bath at 80 ° C overnight, and the reaction was followed by thin layer chromatography until the disappearance of the starting material, the reaction mixture was concentrated, and the residue obtained was purified by the column chromatography to give the title product 5-cyclopentyl- 5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole- 2-Protonamine 16 g (100 mg, colorless oily liquid), yield: 61.3%.

MS m/z (ESI): 4021.

Eighth step

5-cyclopentyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopentyl And [c] Preparation of pyrrole-2-carboxamide tartrate

5-Cyclopentyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-ring Pentacene [c]pyrrole-2-carboxamide 16g (102 mg, 0.254 mmol) was dissolved in 2 mL of ethyl acetate with stirring, and 3 mL of tartaric acid in acetone was added and stirred for 30 minutes. A white solid precipitated and added a small amount. Hexane, stirring was continued, and filtration gave the title product 5-cyclopentyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N, N-Dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide tartrate 16 (88 mg, white solid), yield: 65%.

Example 17

5-benzyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopenta [c]pyrrole-2-carboxamide tartrate

first step

5-benzyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopenta [c]Preparation of pyrrole-2-carboxamide

5-Amino-5-benzyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 14f (0.1 g, 0.35 mmol), 1- (2-Chloro-ethenyl)-2-cyano-pyrrolidine (120.4 mg, 0.7 mmol) and potassium carbonate (49 mg, 0.35 mmol) dissolved in 3 mL of N,N-dimethylformamide with stirring The amine was reacted in an oil bath at 80 ° C overnight, and the reaction was followed by thin layer chromatography until the disappearance of the starting material. The reaction mixture was concentrated, and the residue obtained was purified by silica gel column chromatography to give the title product 5-benzyl-5-[ 2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-yl Indoleamine 17a (56 mg, colorless oily liquid), yield: 40%.

MS m/z (ESI): 42.

Second step

5-benzyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopenta [c]Preparation of pyrrole-2-carboxamide tartrate

5-Benzyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopentyl And [c]pyrrole-2-carboxamide 17a (56 mg, 0.132 mmol) was dissolved in 1 mL of ethyl acetate with stirring, and 1 mL of tartaric acid in acetone was added and stirred for 30 minutes. A white solid precipitated and a small amount of hexane was added. The alkane was stirred and filtered to give the title product 5-benzyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-yloxy-ethylamino]-N,N- Dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide tartrate 17 (50 mg, white solid), yield: 67%.

_{^{1 H NMR (DMSO-D 6}} , 400MHz) δ 7.32-7.19 (m, 5H), 4.78 (m, 1H), 4.23 (s, 2H), 3.8-3.0 (m, 8H), 2.75 (s, 2H) , 2.73 (s, 6H), 2.18 (m, 2H), 2.16 (m, 2H), 1.87 (m, 3H), 1.35 (m, 3H).

Example 18

5-[2-(2-Cyano-4-fluoro-pyrrolidin-1-yl)-2-yloxy-ethylamino]-N,N,5-trimethyl-perhydro-cyclopentyl And [c]pyrrole-2-carboxamide p-toluenesulfonate

first step

5-[2-(2-Cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro-cyclopenta [c]Preparation of pyrrole-2-carboxamide

5-Amino-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 11c (1.3 g, 7.58 mmol), (2S, 4S) under nitrogen atmosphere 1-(2-chloroethenyl)-4-fluoro-2-cyanopyrrolidine 13g (1.74 g, 9.1 mmol), potassium carbonate (1.26 g, 9.1 mmol) and 30 mL of N,N-dimethyl Baseamylamine and 18 mL of dichloromethane were added to the reaction flask, and the reaction was carried out in an oil bath at 30 ° C overnight. The reaction was followed by thin layer chromatography, the starting material disappeared, and N,N-dimethylformamide was removed under reduced pressure. Purification by column chromatography gave the title product 5-[2-(2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5 Trimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 18a (1.39 g, white solid), yield: 50%.

MS m/z (ESI): 348.2 (M + 1).

Second step

5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro-cyclopenta[c]pyrrole Preparation of 2-carbamamine p-toluenesulfonate

5-[2-(2-Cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl- at room temperature Perhydro-cyclopenta[c]pyrrole-2-carboxamide 18a (0.9 g, 2.47 mmol) was dissolved in 5 mL of dichloromethane with stirring, then p-toluenesulfonic acid monohydrate (469 mg, 2.47 mmol) Dissolved in 3 mL of acetone under stirring, and added a solution of p-toluenesulfonic acid in acetone to the above solution. A white precipitate gradually formed during the dropwise addition. After stirring for 30 minutes, 1 mL of n-hexane was added and stirred, and the obtained was obtained. The title product 5-[2-(2-cyano-4-fluoro-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N,5-trimethyl-perhydro- Cyclopenta[c]pyrrole-2-carboxamide p-toluenesulfonate 18 (1.3 g, white solid), yield: 93%.

MS m/z (ESI): 366.1 (M + 1).

_{^{1 H NMR (DMSO-D 6}} , 400MHz) δ 7.74 (d, 2H), 7.29 (d, 2H), 5.57-5.44 (m, 1H), 5.07 (m, 1H), 4.19-4.07 (m, 4H) , 3.49 (m, 2H), 3.35 (m, 2H), 3.0 (m, 2H), 2.9 (s, 6H), 2.6 (m, 2H), 2.39 (m, 5H), 1.57 (m, 2H), 1.34 (s, 3H).

Example 19

5-ethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopenta [c]pyrrole-2-carboxamide tartrate

first step

5-cyano-5-ethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Cyano-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 12a (3.0 g, 14.5 mmol) was dissolved in 60 mL of anhydrous under stirring with nitrogen. Tetrahydrofuran, adding ethyl iodide (4.52 g, 29 mmol) at room temperature, dropwise adding lithium hexamethyldidecylamine (29 mL, 29 mmol), reacting at room temperature for 2 hours, followed by thin layer chromatography After the reaction was completed, the mixture was evaporated to dryness, and ethyl acetate (200 mL×3) was evaporated. Purification by hydrazine column chromatography to give 5-cyano-5-ethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 19a (1.64 g, light yellow oil).

MS m/z (ESI): 236.3 (M + 1).

¹ H NMR (DMSO-D ₆ , 400 MHz) δ 3.5-3.0 (m, 4H), 2.7 (s, 6H), 1.9-1.2 (m, 8H), 1.19 (m, 3H).

Second step

2-Dimethylamine-mercapto-5-ethyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid preparation

Dissolve 5-cyano-5-ethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 19a (1.54 g, 6.55 mmol) in 60 mL two with stirring In methyl chloride, diisobutylaluminum hydride (19.6 mL, 19.6 mmol) was added dropwise at 0 ° C, and the mixture was stirred at 0 ° C for 30 minutes. The reaction was traced by thin layer chromatography until the starting material disappeared. 1.5 mL of water, dried over anhydrous sodium sulfate, filtered, filtered, and then evaporated to give the title compound 5-ethyl-5-carbazyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole 2-carbamamine 19b (0.4 g, pale yellow liquid), yield: 26%.

MS m/z (ESI): 239.1 (M + 1).

third step

Preparation of 5-ethyl-2-dimethylaminomethylindenyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid

Dissolve 5-ethyl-5-methylindenyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 19b (0.4 g, 1.68 mmol) in 18 mL with stirring To a mixed solvent of tetrahydrofuran and 9 mL of water, sodium dihydrogen phosphate dihydrate (787 mg, 5.04 mmol), sodium chlorite (0.454 g, 5.04 mmol) and 2-methyl-2-butene (354 mg, 5.06 mmol), the reaction was carried out at 0 ° C for 2 hours under nitrogen atmosphere, and the reaction was followed by thin layer chromatography until the disappearance of the starting material. The aqueous phase was extracted with ethyl acetate. The organic phase was combined, dried over anhydrous magnesium sulfate and filtered. Concentration by pressure gave the title product 5-ethyl-2-dimethylaminocarbazyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid 19c (0.426 g, pale yellow oily liquid). Rate: 100%, which is used directly for the next reaction.

MS m/z (ESI): 255.2 (M + 1).

the fourth step

Preparation of 5-ethyl-2-dimethylaminomethylmercapto-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide

5-Ethyl-2-dimethylaminomethylindenyl-perhydro-cyclopenta[c]pyrrole-5-carboxylic acid 19c (0.56 g, 2.2 mmol) was dissolved in 60 mL with stirring under ice-cooling In acetone, triethylamine (0.245 mg, 2.43 mmol) was added dropwise at -5 ° C, 2 mL of ethyl chloroformate (263 mg, 2.43 mmol) in acetone was added, stirred for 15 minutes, and 2 mL of sodium azide was added. (0.286 g, 4.4 mmol) of aqueous solution, maintaining the temperature at 0 to -5 ° C, continue the reaction for 30 minutes, trace the reaction by thin layer chromatography until the disappearance of the starting material, add 10 mL of water, with ethyl acetate (10 mL × 5) The organic phase was combined, dried over anhydrous magnesium sulfate, filtered, filtered, evaporated, evaporated, evaporated. Pyrrole-5-carbonyl azide 19d (0.5 g, pale yellow oil) which was used directly in the next step.

the fifth step

Preparation of 5-ethyl-5-isocyanato-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

5-Ethyl-2-dimethylaminocarbazyl-perhydro-cyclopenta[c]pyrrole-5-carbonyl azide 19d (0.5 g, 1.86 mmol) was dissolved in 30 mL of toluene with stirring. After heating under reflux for 2 hours, the reaction was followed by thin layer chromatography until the disappearance of the starting material, and the reaction mixture was evaporated to dryness to give the titled product 5-ethyl-5-isocyano-N,N-dimethyl-all. Hydrogen-cyclopenta[c]pyrrole-2-carboxamide 19e was used directly in the next reaction.

Step 6

Preparation of 5-amino-5-ethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide

12 mL of 8 N hydrochloric acid was added dropwise to the 5-ethyl-5-isocyanato-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2- obtained in the previous step at room temperature. The megamine 19e was stirred for 30 minutes, and the pH was adjusted to 9-10 with 8N sodium hydroxide solution. The aqueous phase was extracted with dichloromethane (30 mL×5). The filtrate was concentrated under reduced pressure, and the obtained residue was purified mjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj -Metformamide 19f (0.3 g, colorless oil), yield: 71%.

MS m/z (ESI): 226.2 (M + 1).

Seventh step

5-ethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopenta [c]Preparation of pyrrole-2-carboxamide

5-Amino-5-ethyl-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 19f (0.202 g, 0.897 mmol), 1- (2-Chloro-ethenyl)-2-cyano-pyrrolidine (185 mg, 1.07 mmol), potassium carbonate (148 mg, 1.07 mmol) and 9 mL dichloromethane/N,N-dimethylformamide A mixed solvent of amine (V/V=1/1) was added to the reaction flask, and the reaction was carried out at 60 ° C for 2 hours. The reaction was traced by thin layer chromatography until the disappearance of the starting material, and the dichloromethane and N in the reaction mixture were removed by concentration under reduced pressure. , N-dimethylformamide, the residue obtained was purified by silica gel column chromatography to give the title product 5-ethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2 -Sideoxy-ethylamino]-N,N-dimethyl-perhydro-cyclopenta[c]pyrrole-2-carboxamide 19 g (120 mg, light yellow oily liquid), yield: 40%.

MS m/z (ESI): 362.2 (M + 1).

Eighth step

5-ethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopenta [c]Preparation of pyrrole-2-carboxamide tartrate

5-Ethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl-perhydro-cyclopentyl And [c]pyrrole-2-carboxamide 19g (120 mg, 0.33 mmol) was dissolved in 1 mL of ethyl acetate with stirring, and 2 mL of tartaric acid (50 mg, 0.33 mmol) in acetone was added dropwise, and stirred for 30 minutes. Filtration gave the title product 5-ethyl-5-[2-(2-cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-N,N-dimethyl- All hydrogen-cyclopenta[c]pyrrole-2-carboxamide tartrate 19 (160 mg, white solid), yield: 94%.

MS m/z (ESI): 362.2 (M + 1).

_{^{1 H NMR (DMSO-D 6}} , 400MHz) δ 4.79 (m, 1H), 4.11 (s, 2H), 3.65 (m, 2H), 3.49 (m, 2H), 3.22 (m, 4H), 2.75 (s , 6H), 2.55 (m, 2H), 2.19 (m, 2H), 2.09 (m, 2H), 1.99 (m, 2H), 1.51 (m, 4H), 0.86 (t, 3H).

Test case Biological evaluation

Determination of inhibitory activity of DPP IV/DPP8/DPP9

The following method was used to determine the inhibition of DPPIV/DPP8/DPP9 enzymatic activity by the compounds of the invention. The ability of the compounds of the invention to inhibit the inhibition of pure DPPIV/DPP8/DPP9 enzyme activity was examined. The inhibition rate or semi-inhibitory concentration IC _{50 of} each compound (the concentration of the compound measured when the enzyme activity is inhibited to 50%) is determined by a fixed amount of the enzyme mixed substrate and different concentrations of the test compound.

Determination of DPP IV inhibitory activity

Materials and Method:

material:

a. White 96-well plate (BMG)

b. Tris Buffer: Prepare 100 mL of 2 mM Tris buffer, dissolve 0.0242 g of Tris in approximately 90 mL of dH ₂ O, adjust pH to 8.00 with HCl and NaOH, and finally add dH ₂ O to 100 mL.

c. DPPIV enzyme (CalBiochem Catalog no. 317630), dissolved in Tris buffer to 2 mM.

d. DPPIV-Glo ^TM by mass (Promega Catalog no.G8350), dissolved in dH ₂ O to 1 mM.

e. DPPIV-Glo. Buffer (Promega Catalog no.G8350)

f. Luciferin detection reagent (Promega Catalog no.G8350)

g. DMSO

h. dH ₂ O

operating:

In the following sequence of operations:

1. Thaw DPPIV-Glo. Buffer before use and equilibrate to room temperature.

2. Buffer frozen luciferin detection reagent before use.

3. Suspension DPPIV-Glo. After adding ultrapure water to the substrate and mixing gently, a 1 mM substrate was prepared.

4. Place the luciferin test reagent in a brown bottle and add DPPIV-Glo. The luciferin detection reagent should be dissolved within 1 minute.

5. Dissolve the test compound in DMSO to 50 times the final working concentration.

6. Add 50 μl of the test compound to 2 μL of each test tube, and add 2 μL of DMSO to the negative control and the blank control.

7. 46 μL of Tris buffer was added to each tube, and 48 μL of Tris buffer was added to the blank.

8. Add 2 μ LDPPIV enzyme to each tube of the negative control and test samples.

9. Vibrate and centrifuge the tube. Transfer all of the material in the tube to a 96-well plate.

10. The ratio of mixed substrate and DPPIV-Glo. was 1:49. Mix the vibrations to mix thoroughly. Allow to stand at room temperature for 30-60 minutes before use.

11. Add 50 μL of DPPIV-Glo. and the substrate mixture to each 96-well plate well and seal the plate with a sealing film.

12. Slowly mix the 96-well material with a plate shaker at 300-500 rpm / 30 s. Incubate for 30 minutes to 3 hours at room temperature.

13. Record the luminescence.

Inhibition rate definition: [1-(S-B)/(N-B)]*100% S: Sample B: blank control N: negative control

Determination of DPP 8 inhibitory activity

Materials and Method:

material:

i. White 96-well plate (BMG)

j. Tris Buffer: Prepare 100 mL of 2 mM Tris buffer, dissolve 0.0242 g of Tris in approximately 90 mL of dH ₂ O, adjust pH to 8.00 with HCl and NaOH, and finally add dH ₂ O to 100 mL.

k. DPP8 enzyme (Bioscience Catalog no. 80080), dissolved in 25 mM Tris-HCl, pH 8.0, 100 mM NaCl, 0.05% Tween-20, 50% glycerol, 3 mM DTT buffer. The assay used a final concentration of 0.1 ng/100 μL assay.

l. DPP8-Glo ^TM by mass (Promega Catalog no.G8350), dissolved in dH ₂ O to 1mM.

m. DPP8-Glo. Buffer (Promega Catalog no.G8350)

n. Luciferin detection reagent (Promega Catalog no.G8350)

o. DMSO

p. dH ₂ O

operating:

In the following sequence of operations:

14. Thaw DPP8-Glo. Buffer before use and equilibrate to room temperature.

15. Buffer frozen luciferin detection reagent before use.

16. Suspension DPP8-Glo. After adding ultrapure water to the substrate and mixing gently, a 1 mM substrate was prepared.

17. Place the luciferin test reagent in a brown bottle and add DPP8-Glo. The luciferin detection reagent should be dissolved within 1 minute.

18. Dissolve the test compound in DMSO to 50 times the final working concentration.

19. Add 50 μl of the test compound to 2 μL of each test tube, and add 2 μL of DMSO to the negative control and the blank control.

20. 46 μL of Tris buffer was added to each tube, and 48 μL of Tris buffer was added to the blank.

21. Add 2 μl of LDPP8 enzyme to each tube of the negative control and test samples.

22. Vibrate and centrifuge the tube. Transfer all of the material in the tube to a 96-well plate.

23. The ratio of mixed substrate and DPP8-Glo. was 1:49. Mix the vibrations to mix thoroughly. Allow to stand at room temperature for 30-60 minutes before use.

24. Add 50 μL of DPP8-Glo. and the substrate mixture to each 96-well plate well and seal the plate with a sealing film.

25. Slowly mix the 96-well material with a plate shaker at 300-500 rpm / 30 s. Incubate for 30 minutes to 3 hours at room temperature.

26. Record the luminescence.

Inhibition rate definition: [1-(S-B)/(N-B)]*100% S: Sample B: blank control N: negative control

Determination of DPP 9 inhibitory activity

Materials and Method:

material:

q. White 96-well plate (BMG)

r. Tris Buffer: Prepare 100 mL of 2 mM Tris buffer, dissolve 0.0242 g of Tris in approximately 90 mL of dH ₂ O, adjust pH to 8.00 with HCl and NaOH, and finally add dH ₂ O to 100 mL.

s. DPP9 enzyme (Bioscience Catalog no. 80090), dissolved in 25 mM Tris-HCl, pH 8.0, 100 mM NaCl, 0.05% Tween-20, 50% glycerol, 3 mM DTT buffer. The reaction was used at a final concentration of 0.01 ng/100 μL assay.

t. DPP9-Glo ^TM by mass (Promega Catalog no.G8350), dissolved in dH ₂ O to 1mM.

u. DPP9-Glo. Buffer (Promega Catalog no.G8350)

v. Luciferin detection reagent (Promega Catalog no.G8350)

w.DMSO

x.dH ₂ O

operating:

In the following sequence of operations:

27. Thaw DPP9-Glo. Buffer before use and equilibrate to room temperature.

28. Buffer frozen luciferin detection reagent before use.

29. Suspension DPP9-Glo. After adding ultrapure water to the substrate and mixing gently, a 1 mM substrate was prepared.

30. Put the luciferin detection reagent into a brown bottle and add DPP9-Glo. The luciferin detection reagent should be dissolved within 1 minute.

31. Dissolve the test compound in DMSO to 50 times the final working concentration.

32. Add 50 μl of the tested compound to each tube 2 μL, negative 2 μL of DMSO was added to the control and blank controls.

33. 46 μL of Tris buffer was added to each tube, and 48 μL of Tris buffer was added to the blank.

34. Add 2 μl of LDPP9 enzyme to each tube of the negative control and test samples.

35. Vibration mix and centrifuge the tube. Transfer all of the material in the tube to a 96-well plate.

36. The ratio of mixed substrate and DPP9-Glo. was 1:49. Mix the vibrations to mix thoroughly. Allow to stand at room temperature for 30-60 minutes before use.

37. Add 50 μL of DPP9-Glo. and the substrate mixture to each 96-well plate well, and seal the plate with a sealing film.

38. The 96-well material was slowly mixed using a plate shaker at 300-500 rpm / 30 s. Incubate for 30 minutes to 3 hours at room temperature.

39. Record the luminescence.

Inhibition rate definition: [1-(S-B)/(N-B)]*100% S: Sample B: blank control N: negative control

Preliminary evaluation of hypoglycemic effect of DPPIV inhibitor

Test purposes:

The effects of DPPIV inhibitor test compound Example 1 and Example 2 on oral glucose tolerance in normal ICR mice were observed, and its hypoglycemic effect in vivo was initially evaluated.

Test animals:

Species, strains: ICR mouse Source: Shanghai Experimental Animal Center, Chinese Academy of Sciences, certificate number: SYXK (Shanghai) 2004-2005 Weight: 25 to 30g Gender: Number of male animals: 40 Feeding conditions: SPF animal house, Temperature: 22-24 ° C, humidity: 45-80%, light: 150-300 Lx, 12 hours day and night alternate.

Test drug:

Name: Example 1 compound batch number: 01 color, form: white powder purity: 96.97% Provided by: Shanghai Hengrui Pharmaceutical Co., Ltd. R & D center preparation method: accurately weigh the drug, add double distilled water, respectively, 0.5, 0.15 And a suspension of 0.05 mg/mL. (Note: Although the compound is shown to be soluble in water on the receiving sample, it is found to be poorly water-soluble in the experiment, and can be dissolved at a low concentration, but there are still visible particles at a concentration of 0.5 mg/mL. We have also tried to formulate this compound with 1% CMC, but the effect is worse than that of double distilled water.) Dosage: Oral administration: 1, 3, 10 mg/kg, volume 20 mL/kg.

Name: Example 2 Compound Lot No.: 01 Color, Form: White Powder Purity: 99.62% Provided by: Shanghai Hengrui Pharmaceutical Co., Ltd. R&D Center Preparation Method: Accurately weigh the drug, add double distilled water to dissolve, mix thoroughly, and prepare A 1.5 mg/mL solution was then diluted to a clear solution of 0.5, 0.15 and 0.05 mg/mL. Dosage: Oral administration: 1, 3, 10 mg/kg, volume 20 mL/kg.

experiment method:

First, the effect of drugs on blood glucose in normal ICR mice

Normal male ICR mice were randomly divided into groups, with 6 rats in each group, which were blank control, positive control group, and different dose administration groups. The specific grouping is as follows:

Test 1:

Blank control: oral double distilled water administration group 1: Oral Example 1 compound 1 mg/kg Oral Example 1 Compound 3 mg/kg Oral Example 1 Compound 10 mg/kg Administration group 2: Oral Example 2 Compound 1 mg/kg orally Example 2 Compound 3 mg/kg Oral Example 2 Compound 10 mg/kg

Trial 2:

Blank control: oral double distilled water administration group 1: Oral Example 1 compound 1 mg/kg Oral Example 1 Compound 3 mg/kg Oral Example 1 Compound 10 mg/kg Administration group 2: Oral Example 2 Compound 1 mg/kg orally Example 2 Compound 3 mg/kg Oral Example 2 Compound 10 mg/kg

After fasting for 6 hours, the animals in each group were given a single test or double distilled water, and oral glucose 2.5 g/kg was given 30 minutes after the administration, 30 minutes before the sugar supply and 30, 60 and 120 minutes after the sugar supply. Blood was taken and serum glucose concentration was measured.

Second, serum glucose determination method:

Use glucose kit to determine the glucose content in the serum, take 250 μl of enzyme working solution, add 5 μl of serum, set up a blank tube (add 5 μl of double distilled water) and standard tube (add 5 μl of glucose standard solution), mix, The water bath was incubated at 37 ° C for 20 minutes, and the blank tube was adjusted to zero, and the color was measured at OD 505 nm.

Serum glucose content BG (mmol / l) = OD _{sample tube} / OD _{standard tube} × 5.55

Data processing and statistical analysis:

1. Statistical analysis of data using mean ± SD and Student - t test

2. Calculate the percentage of blood glucose drop at 30 minutes after feeding sugar and the area under the curve AUC

test results:

Test 1:

Male ICR mice were orally administered with double distilled water, different doses of the test article, Example 1, and Example 2, after 6 hours of fasting, and each group was given an oral glucose tolerance test after 30 minutes of administration. The results showed that the serum glucose concentration of the blank control group was significantly increased after oral administration of 2.5 g/kg of glucose, and reached a peak at 30 minutes. The blood glucose of the mice of the low, medium and high doses of the compound of Example 1 was significantly lower than that of the blank control group at 30 minutes, and the percentages of blood glucose decreased were 19.16%, 22.85 and 31.85%, respectively. The blood glucose of the mice in each dose group of Example 2 was significantly lower than that of the blank control group at 30 minutes after the administration of sugar (P<0.01), and decreased by 25.54%, 25.92 and 26.93%, respectively, compared with the blank control group.

Trial 2:

Male ICR mice were given double distilled water and different doses after fasting for 6 hours. The compound of Example 1 and the compound of Example 2 were administered to each group for 30 minutes after administration for oral glucose tolerance test. The results showed that the serum glucose concentration of the mice in the blank control group was significantly increased after oral administration of 2.5 g/kg of glucose, and reached the peak at 30 minutes. The blood glucose of the mice in each dose of SHR1039 was significantly lower than that of the blank control group at 30 minutes after the administration of sugar (P< 0.01), compared with the blank control group, decreased by 26.10%, 30.24 and 32.05%, respectively. The blood glucose of the SHR1040 low, medium and high dose groups was significantly lower than that of the blank control group at 30 minutes, and the percentage of blood glucose decreased by 24.51%, 26.96% and 27.75%, respectively.

in conclusion:

The results of the two tests in this report showed that the compound of Example 1 and the compound of Example 2 had significant hypoglycemic effects in the oral glucose tolerance test in normal ICR mice. Among them, the compound of Example 1 tested showed a good dose-effect relationship.

Effect of DPPIV inhibitory compounds on oral glucose tolerance in KKAy mice

The effects of the compound of Example 1 and the compound of Example 2 on oral glucose tolerance in type 2 diabetic KKAy mice were observed, and the hypoglycemic effect in vivo was initially evaluated.

Test animals:

Species, strains: KKAy mouse Source: Shanghai Experimental Animal Center, Chinese Academy of Sciences, Certificate No.: SYXK (Shanghai) 2004-2005 Weight: 40 to 55g Gender: Female: 52; Male: 33 Feeding conditions: SPF animal house, temperature: 22-24 ° C, humidity: 45-80%, light: 150-300 Lx, 12 hours day and night alternate.

Test drug:

Name: Compound of Example 1 and compound of Example 2 Preparation method: Accurately weigh the drug, dissolve it in double distilled water, mix well, prepare a suspension of 3 mg/mL, and then dilute to 1, 0.3 and 0.1 mg/mL. Transparent solution. Dosage: Oral administration: 1, 3, 10, 30 mg/kg, volume 10 mL/kg.

experiment method:

Effect of test substance on blood glucose of KKAy mice

Normal KKAy mice were divided into body weight and fasting blood glucose after 6 hours of fasting, and 5 rats in each group were blank control and different dose administration groups. The specific experiment is as follows:

Test 1: Male 0704 blank control: Oral double distilled water SHR1039 group: Oral Example 1 Compound 10 mg/kg Oral Example 1 Compound 30 mg/kg Test 2: Female 0816 blank control: Oral double distilled water SHR1039 group: Oral Example 1 Compound 3 mg/kg Oral Example 1 Compound 10 mg/kg Trial 3: Male 0712 Blank control: Oral double distilled water SHR1040 group: Oral Example 2 Compound 3 mg/kg Oral Example 2 Compound 10 mg/kg Test 4: Female 0907 blank control: Oral double distilled water SHR1040 group: Oral Example 2 compound 3 mg/kg orally Example 2 Compound 10 mg/kg

After 6 hours of fasting, the animals in each group were given a single test or double distilled water, and oral glucose 2.5 g/kg (female KKAy mice) or 1.5 g/kg (male KKAy mice) 30 minutes after the administration. The serum glucose concentration was measured with a blood glucose meter at 0, 30, 60, and 120 minutes after the sugar administration.

Data processing and statistical analysis:

3. Statistical analysis of data using mean ± SD and Student - t test or Anova

4. Calculate the percentage of blood glucose drop at 30 minutes after giving sugar and the area under the curve AUC

test results:

Compound Example 1: Test 1, 2

Male KKAy mice were orally administered with double distilled water and different doses of the test compound of Example 1 after 6 hours of fasting. Each group was given an oral glucose tolerance test after 30 minutes of administration. The results showed that the serum glucose concentration of the blank control group was significantly increased after oral administration of 1.5 g/kg of glucose, and peaked at 30 minutes. The blood glucose ratio of the mice in the 10 and 30 mg/kg groups at 30 minutes after the administration of sugar in Example 1 The blank control group decreased, which was 16.22% and 17.15% lower than the blank control group, respectively.

Female KKAy mice were orally administered with double distilled water and different doses of the test compound of Example 1 after 6 hours of fasting. Each group was given an oral glucose tolerance test after 30 minutes of administration. The results showed that the serum glucose concentration of the blank control group was significantly increased after oral administration of 2.5 g/kg of glucose, and peaked at 30 minutes. The blood glucose and blank control of the mice in the 3 and 10 mg/kg groups were given 30 minutes after the glucose administration. Compared with the group, the decrease rate was 40.63% and 24.68%.

V. Compound Example 2: Test 3, 4

Male KKAy mice were orally administered with double distilled water and different doses of the test compound of Example 2 after 6 hours of fasting. Each group was given an oral glucose tolerance test after 30 minutes of administration. The results showed that the serum glucose concentration of the blank control group was significantly increased after oral administration of 1.5 g/kg of glucose, and peaked at 30 minutes. The blood glucose of the mice in the 10 and 30 mg/kg groups at 30 minutes after glucose administration was compared with the blank control. The group decreased, compared with the blank control group, decreased by 13.79% and 12.23%, respectively.

Female KKAy mice were orally administered with double distilled water and different doses of the test compound of Example 2 after 6 hours of fasting. Each group was given an oral glucose tolerance test after 30 minutes of administration. The results showed that the serum glucose concentration of the mice in the blank control group was significantly increased after oral administration of 2.5 g/kg of glucose, and peaked at 30 minutes. The blood glucose of the mice in the 10 mg/kg group of Example 2 was 30 minutes after the glucose administration. The decrease (P=0.075, anova) decreased by 21.55%, but there was no significant difference due to individual differences.

in conclusion:

The compound of Example 1 and the compound of Example 2 had a certain hypoglycemic effect in the oral glucose tolerance test of type 2 diabetes KKAy mice.

Claims (21)

A compound represented by the formula (IA): or a pharmaceutically acceptable salt thereof: Where: R is R _{1 is} selected from -C(O)NR ₃ R ₄ , -C(O)R ₃ or -C(O)OR ₃ ; R _{2 is} selected from a hydrogen atom or an alkyl group, wherein the alkyl group is optionally further Substituted with one or more substituents selected from cycloalkyl or aryl; R ₃ and R ₄ are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group, a heteroaryl group or a heterocycloalkyl group, wherein The isoalkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl group is further optionally further selected from one or more selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy Substituents for aryl, aryloxy, heteroaryloxy, halogen, hydroxy, amine, cyano, hydroxyalkyl, heterocycloalkyl, heterocycloalkoxy, trifluoromethyl, carboxy or carboxylate groups Substituting; or, R ₃ and R ₄ together with the N atom form a 3 to 8 membered heterocyclic group, wherein the 5 to 8 membered heterocyclic ring optionally further contains one or more N, O or S atoms, and The 3- to 8-membered heterocyclic ring is optionally further further selected from one or more selected from the group consisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkoxy, aryloxy, hydroxy Alkyl, heterocycloalkyl, carboxyl, carboxylate or halogen Substituent group; R ₅ is selected from hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl, wherein such alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl Further optionally further selected from one or more selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy, heteroaryloxy, halogen, hydroxy, amine, Substituted with a substituent of an alkylamino group, a cyano group, a hydroxyalkyl group, a heterocycloalkyl group, a heterocycloalkoxy group, a carboxyl group or a carboxylate group; R ₆ and R ₇ are each selected from an alkyl group, an aryl group, and a heteroaryl group. , haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkynyl, alkoxy, aryloxy, hydroxyalkyl, heterocycloalkyl, carboxy, carboxylate or halogen; W is carbon, sulfur Or an oxygen atom; when W is carbon, it may be further substituted by R ₆ or R ₇ . A compound according to claim 1 or a pharmaceutically acceptable salt thereof, which comprises a compound represented by the following formula (IB) or a pharmaceutically acceptable salt thereof: Where: R is R _{1 is} selected from -C(O)NR ₃ R ₄ , -C(O)R ₃ or -C(O)OR ₃ ; R _{2 is} selected from a hydrogen atom or an alkyl group, wherein the alkyl group is optionally further subjected to a Or a plurality of substituents selected from a cycloalkyl group or an aryl group; R ₃ and R ₄ are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group, a heteroaryl group or a heterocycloalkyl group, wherein The alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl group is further optionally further selected from one or more selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy Substituent substitution of aryloxy, heteroaryloxy, halogen, hydroxy, amine, cyano, hydroxyalkyl, heterocycloalkyl, heterocycloalkoxy, trifluoromethyl, carboxy or carboxylate groups Or, R ₃ and R ₄ together with the N atom form a 3 to 8 membered heterocyclic group, wherein the 5 to 8 membered heterocyclic ring optionally further contains one or more N, O or S atoms, and the 3 The 8-membered heterocyclic ring is optionally further further selected from one or more selected from the group consisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkoxy, aryloxy, hydroxyalkane Substituent, heterocycloalkyl, carboxyl, carboxylate or halogen substitution Substituted; R ₅ is selected from hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl, wherein such alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl Optionally further selected from one or more selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy, heteroaryloxy, halogen, hydroxy, amine, alkane Substituted with a substituent of an amine group, a cyano group, a hydroxyalkyl group, a heterocycloalkyl group, a heterocycloalkoxy group, a carboxyl group or a carboxylate group; and R ₆ and R ₇ are each selected from an alkyl group, an aryl group, a heteroaryl group, Haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkynyl, alkoxy, aryloxy, hydroxyalkyl, heterocycloalkyl, carboxy, carboxylate or halogen; W is carbon, sulfur or An oxygen atom; when W is carbon, it may be further substituted with R ₆ or R ₇ . A compound according to claim 1 or a pharmaceutically acceptable salt thereof, which comprises a compound represented by the following formula (IC) or a pharmaceutically acceptable salt thereof: Where: R is R _{1 is} selected from -C(O)NR ₃ R ₄ , -C(O)R ₃ or -C(O)OR ₃ ; R _{2 is} selected from a hydrogen atom or an alkyl group, wherein the alkyl group is optionally further subjected to a Or a plurality of substituents selected from a cycloalkyl group or an aryl group; R ₃ and R ₄ are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, an aryl group, a heteroaryl group or a heterocycloalkyl group, wherein The alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl group is further optionally further selected from one or more selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy Substituent substitution of aryloxy, heteroaryloxy, halogen, hydroxy, amine, cyano, hydroxyalkyl, heterocycloalkyl, heterocycloalkoxy, trifluoromethyl, carboxy or carboxylate groups Or, R ₃ and R ₄ together with the N atom form a 3 to 8 membered heterocyclic group, wherein the 5 to 8 membered heterocyclic ring optionally further contains one or more N, O or S atoms, and the 3 to The 8-membered heterocyclic ring is optionally further further selected from one or more selected from the group consisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy, hydroxy, amine, cyano, alkoxy, aryloxy, hydroxyalkyl a heterocyclic alkyl group, a carboxyl group, a carboxylate group or a halogen substituent ; R ₅ is selected from hydrogen, alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl, wherein such alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl any Further selected from one or more selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy, heteroaryloxy, halogen, hydroxy, amine, alkylamine Substituted with a substituent of a cyano group, a cyano group, a hydroxyalkyl group, a heterocycloalkyl group, a heterocycloalkoxy group, a carboxyl group or a carboxylate group; and R ₆ and R ₇ are each selected from the group consisting of an alkyl group, an aryl group, a heteroaryl group, and an alkyl halide. , haloalkoxy, hydroxy, amine, cyano, alkynyl, alkoxy, aryloxy, hydroxyalkyl, heterocycloalkyl, carboxyl, carboxylate or halogen; W is carbon, sulfur or oxygen Atom; when W is carbon, it may be further substituted by R ₆ or R ₇ . The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the salt is a salt of a compound of the formula (I) with an acid selected from the group consisting of hydrochloric acid, p-toluenesulfonic acid, tartaric acid, and Malay. Acid, lactic acid, methanesulfonic acid, sulfuric acid, phosphoric acid, citric acid, acetic acid or trifluoroacetic acid. The compound of claim 4 or a pharmaceutically acceptable salt thereof, wherein the acid is p-toluenesulfonic acid, hydrochloric acid, tartaric acid or trifluoroacetic acid. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, which comprises a compound having the following structure: A compound according to claim 1 or a pharmaceutically acceptable salt thereof, which comprises a compound having the following structure: A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. A compound of the following formula (ID) which is an intermediate for the synthesis of a compound of the formula (I) as in claim 1 of the patent application: Wherein R _{1 is} selected from -C(O)NR ₃ R ₄ , -C(O)R ₃ or -C(O)OR ₃ ; and R ₃ and R ₄ are each selected from a hydrogen atom, an alkyl group, a cycloalkyl group, Aryl, heteroaryl or heterocycloalkyl, wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl group is further further selected from one or more selected from the group consisting of alkyl, cycloalkyl , aryl, heteroaryl, alkoxy, cycloalkoxy, aryloxy, heteroaryloxy, halogen, hydroxy, amine, cyano, hydroxyalkyl, heterocycloalkyl, heterocycloalkoxy Substituting a substituent of a trifluoromethyl group, a carboxyl group or a carboxylate group; or, R ₃ and R ₄ together with the N atom form a heterocyclic group of 3 to 8 members, wherein the 5- to 8-membered heterocyclic ring is optionally further Containing one or more N, O or S atoms, and the 3 to 8 membered heterocyclic ring is optionally further further selected from one or more selected from the group consisting of alkyl, aryl, heteroaryl, haloalkyl, haloalkoxy, hydroxy, amine Substituent substitution of a cyano group, a cyano group, an alkoxy group, an aryloxy group, a hydroxyalkyl group, a heterocycloalkyl group, a carboxyl group, a carboxylate group or a halogen. A method of preparing an intermediate (ID) according to claim 9 of the patent application, the method comprising: 5-sided oxy-perhydro-cyclopenta[c]pyrrole-2-carboxylic acid tert-butyl ester was reacted with trifluoroacetic acid (TFA) in a solvent under ice bath to obtain a perhydro-cyclopenta[ c] pyrrol-5-one trifluoroacetate; The perhydro-cyclopenta[c]pyrrole-5-one trifluoroacetate is reacted with hydrazine chloride or an ester in the presence of a base to give a compound of the formula (ID); wherein R ₁ is as defined in the patent application 9 items mentioned. The method of claim 10, wherein the solvent is dichloromethane. A method of preparing a compound of the formula (IB) according to claim 2 of the patent application, the method comprising: The intermediate (ID) is reacted with a different amine, substituted sodium borohydride in a solvent under basic conditions at room temperature to give a compound of the formula (IB); wherein R, R ₁ , R ₂ are as defined above The scope of patent application is mentioned in item 2. The method of claim 12, wherein the solvent is methanol or ethanol. A method of preparing a compound of the general formula (IC) according to claim 3 of the patent application, the method comprising: After heating the toluene solution of potassium t-butoxide and methyltriphenylphosphonium iodide, the intermediate (ID) compound is added to the above solution at room temperature to obtain a methylene azabicyclo compound; The methylene azabicyclo compound is reacted with trimethyl decanoyl at room temperature in the presence of silver perchlorate in a solvent to obtain an isocyanoazabicyclo compound; The isocyanoazabicyclo compound is reacted with an acid at room temperature in a solvent to obtain an amine azabicyclo compound; The amino azabicyclo compound is reacted with a halide in a solvent to obtain a compound of the formula (IC); wherein R, R ₁ , R ₂ are as defined in claim 3 of the scope of the patent application. The method of claim 14, wherein the solvent used for the reaction of the methylene azabicyclo compound with the trimethylcyanamide is dichloromethane; the isocyanoazabicyclo compound and the acid The solvent used in the reaction is ethanol and the acid is hydrochloric acid. A method of preparing a compound of the formula (IB) according to claim 2 of the patent application, the method comprising: The intermediate (ID) compound and p-toluenesulfonylmethyl isocyanide are cyanated in a solvent to obtain a cyanoazabicyclo compound; The cyanoazabicyclo compound is reacted with a halide in a solvent in the presence of lithium hexamethyldidecylalkylamine to obtain an R ₂ -substituted cyanoazabicyclo compound; The R ₂ -substituted cyanoazabicyclo compound is hydrolyzed by an acid to obtain an R ₂ -substituted carboxyazabicyclo compound; or the R ₂ -substituted cyanoazabicyclo compound is solvent Under the ice bath, it is reacted with reducing agent hydrogenated diisobutylaluminum (DIBAL-H). The cyano group is first reduced to an aldehyde group, and then the obtained aldehyde is mixed in a tetrahydrofuran/water mixed solvent with sodium dihydrogen phosphate in an ice bath. , sodium chlorite and 2-methyl-2-butene are reacted to obtain a R ₂ -substituted carboxyazabicyclo compound; The R ₂ -substituted carboxyazabicyclo compound is reacted with an alkali metal azide in the presence of an alkaline condition and ethyl chloroformate to obtain an R ₂ -substituted azidocarbonyl azabicyclo compound; The R ₂ -substituted azidocarbonylcarbonyl azabicyclo compound is heated in a solvent, stirred in an acidic solution, and then adjusted to a basic pH to obtain an R ₂ -substituted amino azabicyclo compound; The R ₂ -substituted amino azabicyclo compound is reacted with a halide in a solvent to obtain a compound of the formula (IB); wherein R, R ₁ , R ₂ are as defined in the second item of the patent application. A method of preparing a compound of the general formula (IC) according to claim 3 of the patent application, the method comprising: The intermediate (ID) is reacted with a reducing agent in a solvent under ice bath to obtain a hydroxyazabicyclo compound; The hydroxyazabicyclo compound is reacted with an alkaline reagent or methanesulfonyl chloride in a solvent under ice to obtain a methanesulfonyl azabicyclo compound; The methanesulfonyl azabicyclo compound is heated and reacted with potassium phthalimide in a solvent to obtain an azabibicyclic compound substituted with phthalimide; The phthalimide substituted azabicyclo compound is heated in a solvent with hydrazine to obtain an amine azabicyclo compound; The amine-substituted azabicyclo compound is heated in a solvent with a halide to give a compound of the formula (IC); wherein R, R ₁ , R ₂ are as defined in claim 3 of the scope of the patent application. The method of claim 17, wherein the reducing agent is lithium tris(t-butoxy)aluminum hydride. The method of any one of claims 11 to 18, which further comprises purifying the obtained compound of the formula (IB) or (IC) in methanol, dichloromethane or ethyl acetate with an acid The reaction is carried out to obtain an acid addition salt thereof. A use of a compound of any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof for the preparation of a dipeptidyl peptidase inhibitor. A use of the composition of claim 8 in the preparation of a dipeptidyl peptidase inhibitor.