Enterolytic co-drugs, their preparation and use

Abstract

The present invention relates to enterically degradable codrugs, their preparation and uses, and in particular, the present invention provides codrug compounds of Formula I. The present invention also provides methods of using such compounds to treat gastrointestinal autoimmune diseases, inflammatory diseases and cancer, as well as methods and intermediates for preparing such compounds.

Description

Technical Field

[0001] The present invention relates to an enteric-degradable co-drug compound. The present invention also relates to a pharmaceutical composition containing such a compound, a method for treating a digestive autoimmune disease, an inflammatory disease and cancer using such a compound, and a method and an intermediate for preparing such a compound.

Background Art

[0002] JAK family (JAK1, JAK2, JAK3, TYK2) inhibitors such as tofacitinib have been approved as therapeutic agents for certain patients suffering from moderate to severe active rheumatoid arthritis (RA) and moderate to severe ulcerative colitis (UC). In many clinical trials of JAK family inhibitors, a large number of adverse events mediated by systemic drug exposure, including serious infections, opportunistic infections and laboratory abnormalities such as lymphopenia, neutropenia, elevated liver enzymes, elevated lipids and elevated serum creatinine, have been reported. All currently marketed JAK inhibitors carry a black box warning for safety risks, including serious infections, malignancies and thrombosis risks. Therefore, in order to develop a safer next-generation JAK family inhibitor drug, it is necessary to limit the systemic exposure amount when treating local inflammatory diseases. For example, when treating UC, while increasing the distribution amount of the JAK family inhibitor in the digestive tract, the systemic exposure amount of the drug should be minimized.

[0003] Berberine is an isoquinoline alkaloid extracted from plants such as Coptis japonica. Berberine is a very safe drug and has a history of use in traditional Chinese medicine for over a thousand years. Its bioavailability is low. Clinically, it is mainly used to treat gastrointestinal diseases such as diarrhea and intestinal infections. Recent studies have shown that berberine also has some therapeutic effects on cardiovascular diseases and the regulation of glucose and lipid metabolism. Berberubine is the main metabolite of berberine in the body. Animal model studies have shown that berberubine has a similar therapeutic effect to berberine on ulcerative colitis. However, currently, there is no effective method in this field to enhance the therapeutic effect of berberubine or its analogues.

[0004] Chronic enteritis mainly includes two types: ulcerative colitis and Crohn's disease. These chronic inflammatory bowel diseases have a long course, are characterized by frequent and recurring attacks, and prolonged inflammation is prone to becoming cancerous. In recent years, the incidence of chronic enteritis has been on the rise. Currently, it is thought that genetics, environment, immunity, and microorganisms may be involved in the development of chronic enteritis, but the exact mechanism has not been clarified. Clinical treatment mainly involves aminosalicylic acid drugs, adrenal glucocorticoid drugs, and immunosuppressants, but certain adverse reactions such as gastrointestinal discomfort and allergic reactions occur. In previous studies, the inventors have discovered that the combination of a JAK inhibitor and a berberine analog can achieve a synergistic effect and superior therapeutic effect for gastrointestinal inflammatory diseases. The present invention aims to design a co-pharmacological compound of an enterogradable berberine analog and a JAK inhibitor so that the two drug molecules are released and enriched in the digestive tract, while systemic exposure of the compound is limited. The synergistic action of the two drug molecules achieves superior therapeutic efficacy and safety. [Overview of the project] [Problems that the invention aims to solve]

[0005] In one embodiment, the present invention provides a co-agent in which a berberine analog (preferably berberbine) and a second therapeutic agent, a JAK family inhibitor (preferably tofacitinib, upadacitinib, or SHR0302), are linked via a degradable covalent bond, designed to increase the content of the JAK family inhibitor and the first therapeutic agent at the site of inflammation in the gastrointestinal tract while minimizing systemic exposure, by targeting and releasing the JAK family inhibitor and the first therapeutic agent in the gastrointestinal tract. [Means for solving the problem]

[0006] A first aspect of the present invention provides a co-pharmaceutical compound represented by the following formula I, formed by coupling a first drug molecule, a second drug molecule, and a linker precursor. [ka] During the ceremony, D1 is the first drug group, and the first drug group is a structural fragment of the first drug molecule that can be linked to the linker (i.e., a fragment obtained by coupling or condensing the first drug molecule with a precursor of the linker and removing the active functional group, and the fragment does not contain the linker portion). D2 is a second drug group, the second drug group is a structural fragment of the second drug molecule that can be linked to the linker, and the first drug molecule and the second drug molecule are synergistic drug molecules (i.e., the second drug molecule is a fragment formed by coupling or condensing with a precursor of the linker, thereby removing the active functional group, and the fragment does not contain the linker portion). Here, the linkage may be a covalent bond that has lost one hydrogen atom, or a covalent bond with a linker by other means, such as a covalent bond formed by a condensation reaction of active groups such as hydroxyl, carboxyl, or amino. Furthermore, the linker has a structure selected from the following group (a), (b), or (c), in each formula, where J1 is linked to the first drug group and J2 is linked to the second drug group. [ka] The Glu has a structure selected from the following group:

Chemical formula

Chemical formula

Chemical formula

[0007] In another preferred example, the linkage includes the drug molecule losing a structural fragment to form a linkage site, or linkage by coordination bonds.

[0008] In another preferred example, in the above formulas (a), (b), and (c), J1 and J2 are each independently -(Y) z-and Y is selected from the group consisting of -NH-, -C(O)-, -C(O)O-, -NHC(O)NH-, -CH=CH-, -NH(CH2)-, -NHC(O)-, -CH2-, -OCH2CH2O-, -O-, -S-, -P(O)2O-, -S(O)2-, -S(O)-, -C(O)NH-, -N=N-, and Y may be substituted with one or more Rs as long as each Y jointly constitutes a chemically stable structure. As long as each L1, L2, L3, L4, L5, L6, and L7 forms a stable divalent group, each L1, L2, L3, L4, L5, L6, and L7 independently forms a C1-C8 alkylene, C 1~6 Alkylene-OC 1~4 Alkylene (-CH2-O-CH2-), C 2~6 Alkenil, C 2~6 Alkinyl, C 3~6 The group consisting of cycloalkyl, C6-C10 arylene, heteroarylene with 5-10 atoms, and heterocyclylene with 3-12 atoms, or -NH-, -C(O)-, -CH=CH-, -NH(CH2)-, -NHC(O)-, -CH2-, -OCH2CH2O-, -O-, -S-, -P(O)2O-, -S(O)2-, -S(O)-, -C(O)NH-, -C(O)O-, -NHC(O)NH-, -N=N-, -C(O)NH(CH2) (1~4) Selected from the group consisting of -NHC(O)-, Furthermore, as long as each Y, L1, L2, L3, L4, L5, L6, and L7 jointly constitutes a chemically stable structure, Y, L1, L2, L3, L4, L5, L6, and L7 may be arbitrarily substituted with one or more R, and the R may be H, -OH, C1-C4 alkyl, halogen, cyano, nitro, -OR4, or C 1~6 Selected from the group consisting of haloalkyl, sulfonic acid group, C0-C4 alkyl-S(O)2-C1~C4 alkyl, formyl, carboxyl, and -COOR4. Furthermore, m, n, p, q, r, s, and t are each independently selected from 0, 1, 2, 3, 4, 5, 6, 7, and 8. z is selected from the group consisting of 0, 1, 2, 3, 4, 5, and 6, and preferably z is selected from 1, 2, and 3.

[0009] In another preferred example, in formulas (a), (b), and (c) above, J1 and J2 are each independently selected from the group consisting of -NH-, -C(O)-, -C(O)O-, -NHC(O)NH-, -CH=CH-, -NH(CH2)-, -NHC(O)-, -CH2-, -OCH2CH2O-, -O-, -S-, -P(O)2O-, -S(O)2-, -S(O)-, -C(O)NH-, and -N=N-.

[0010] In another preferred example, J1 and J2 are independently methylene, [ka] It is selected from the group consisting of the following.

[0011] In another preferred example, J1 is methylene, [ka] It is selected from the group consisting of the following.

[0012] In another preferred example, J2 is methylene, [ka] It is selected from the group consisting of the following.

[0013] In another preferred example, the first drug molecule is berberine, berberbine, and its analogues. In another preferred example, the second drug molecule is a JAK family inhibitor and its analogues.

[0014] In another preferred example, the linker has the structure shown below. [ka]

[0015] In another preferred example, the first drug molecule is a drug molecule of formula II, formula III, or formula IV below, [ka] During the ceremony, Ro, Rp, Rq, Rr, Rs, and Rt are each independently selected from the group consisting of H, substituted or unsubstituted C1-C4 alkyl groups, and substituted or unsubstituted C1-C4 alkoxy groups, or Ro, Rp, Rq, Rr, Rs, and Rt, when located on two adjacent atoms, form a 5-7 membered heterocycle together with the atom linked to them, where substitution means that the H atom on the group is substituted with one or more substituents selected from the group consisting of halogens, C1-C4 alkyl groups, and phenyl groups.

[0016] In another preferred example, the JAK family inhibitors and their analogues include tofacitinib, ruxolitinib, oclacitinib, baricitinib, peficitinib, abrocitinib, filgotinib, upadacitinib, delgocitinib, itacitinib, fedratinib, and desernotinib. The group consisting of (Decernotinib), SHR-0302, AZD-4205, ASN-002, BMS-986165, PF-06700841, PF-06651600, R-348, INCB-52793, ATI-501, ATI-502, NS-018, KL-130008, or a deuterated derivative of the above molecules is selected.

[0017] In another preferred example, the first drug group is [ka] Selected from the group consisting of, Alternatively, the first drug group is a group formed when a drug molecule selected from the following group loses one hydrogen atom. [ka]

[0018] In another preferred example, the first drug group has a structure represented by the following formula. [ka]

[0019] In another preferred example, the second drug group is selected from the following group. [ka] [ka]

[0020] In another preferred example, the second drug group is selected from the following group. [ka]

[0021] In another preferred example, the first drug group is [ka] The second drug group is [ka] That is the case.

[0022] In another preferred example, A-(L7) p -J2- has the structure shown by the following formula. [ka]

[0023] In another preferred example, the -A(Glu)-(L7) p -J2- has a structure selected from the following group. [ka]

[0024] In another preferred example, the above-(L1) m -and-(L2) n -Each of these independently has a structure selected from the following group, [ka] In each of the above formulas, Ra, Rb, and Rc are each independently groups formed by the loss of one hydrogen atom from an amino acid selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosine, aspartate, histidine, asparagine, glutamate, lysine, glutamine, methionine, arginine, serine, threonine, cysteine, and proline.

[0025] In another preferred example, the linker is selected from the following groups (A), (B), or (C): (A) Group is -L a -L- has the structure, the L a The structure is selected from the following group: [ka] Furthermore, L has the structure shown below, where * represents L and L a This is the connection point. [ka]

[0026] Group (B): [ka]

[0027] Group (C): [ka]

[0028] In another preferred example, the compound is selected from the following group. [ka] [ka] [ka] [ka] [ka] [ka]

[0029] In another preferred example, the compound is selected from the following group. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0030] A second aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the compound described in the first aspect of the present invention or its stereoisomer or racemate or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. In another preferred example, the pharmaceutical composition is an enteric-coated preparation.

[0031] In another preferred example, the pharmaceutical composition is used to treat a disease selected from the group consisting of gastrointestinal inflammatory diseases (e.g., ulcerative colitis, Crohn's disease, colitis associated with immune checkpoint inhibitor therapy, collagen colitis, lymphocytic colitis, pouchitis, acute / chronic gastritis, acute / chronic appendicitis), gastroenteritis due to radiotherapy or chemotherapy, autoimmune diseases of the digestive system (e.g., graft-versus-host disease, sprue, autoimmune bowel disease), peptic ulcers, irritable bowel syndrome, gastric cancer, esophageal cancer, and colon cancer.

[0032] A third aspect of the present invention provides the use of a precursor compound described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described in the second aspect of the present invention, for use in the prevention and treatment of digestive system disorders.

[0033] In another preferred example, the digestive dysfunction is a gastrointestinal inflammatory disease. In another preferred example, the gastrointestinal inflammatory disease is selected from the group consisting of ulcerative colitis, Crohn's disease, and colitis associated with immune checkpoint inhibitor therapy.

[0034] Within the scope of the present invention, the technical features described above and the technical features specifically described below (for example, in the examples) may be combined to constitute a novel or preferred technical embodiment. Due to space limitations, a detailed explanation will not be provided here. [Brief explanation of the drawing]

[0035] [Figure 1] The curves show the changes in the concentration of the compound from Example 8 over time in various tissues when the compound from Example 8 was orally administered to mice. [Figure 2]The curves show the time-dependent changes in the concentration of berberbine in various tissues when the compound of Example 8 was orally administered to mice. [Figure 3] The curves show the time-dependent changes in the concentration of tofacitinib in various tissues after oral administration of the compound of Example 8 to mice. [Figure 4] The AUC0-24h of the content of berberbine in various tissues after oral administration of the compounds of Examples 1 and 8 to mice is shown. [Figure 5] The AUC0-24h of tofacitinib content in various tissues after oral administration of the compounds of Examples 1 and 8 to mice is shown. [Figure 6] This diagram shows the change in disease index when the compound from Example 8 was administered to mice in an oxazolone enema model. [Modes for carrying out the invention]

[0036] Through long-term and in-depth research, the inventors discovered that by preparing and applying a co-drug formulation of a JAK family inhibitor and a berberine analog, superior therapeutic effects were obtained for the treatment of gastrointestinal diseases at the same dose compared to monotherapy. Furthermore, the co-drug molecule design allowed for targeted drug release. Based on these findings, the inventors completed the present invention.

[0037] term Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art.

[0038] As used herein, the terms “contain” or “include” may be open, semi-closed, or closed. In other words, the terms also include “substantially composed of” or “composed of.”

[0039] In this application, the term "alkyl" means a fully saturated linear or branched hydrocarbon chain group, either as a group or as part of another group, consisting only of carbon and hydrogen atoms, having, for example, 1 to 12 (preferably 1 to 8), more preferably 1 to 6 carbon atoms, and including, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, heptyl, 2-methylhexyl, 3-methylhexyl, octyl, nonyl, and decyl, and linked to the rest of the molecule by a single bond. In relation to the present invention, the term "C1-C6 alkyl" means an alkyl group having 1 to 6 carbon atoms.

[0040] In this application, the term "6-10 membered aromatic ring" means an aromatic ring having 6 to 10 ring atoms, which are carbon atoms, as a group or as part of another group. The aromatic ring may be monocyclic or dicyclic. Examples include benzene rings, naphthalene rings, and other similar groups.

[0041] In this application, the term "5-10 membered heteroaromatic ring" means a heteroaromatic ring having 5 to 10 ring atoms, at least one (which may be 1, 2, or 3) of which are heteroatoms selected from nitrogen, oxygen, and sulfur, as part of a group or other group. The heteroaromatic ring may be monocyclic or dicyclic. Examples include pyriminopyrazole rings, pyrazinoimidazole rings, pyridinopyrazole rings, pyridinoimidazole rings, pyridinopyrimidine rings, and pyridinopyridine rings.

[0042] In this application, the term “heterocyclyl” means a stable 3-20 membered non-aromatic cyclic group comprising 3-14 carbon atoms and 1-6 heteroatoms selected from nitrogen, phosphorus, oxygen, and sulfur, either as a group or as part of another group. Unless otherwise specified herein, heterocyclyls can be monocyclic, dicyclic, tricyclic, or more than monocyclic ring systems, including fused ring systems, bridging ring systems, or spirocyclic systems, wherein the nitrogen, carbon, or sulfur atoms in the heterocyclyl may optionally be oxidized, the nitrogen atoms may optionally be quaternized, and the heterocyclyl may be partially or completely saturated. Heterocyclyls can be linked to the rest of the molecule by single bonds via carbon atoms or heteroatoms. Among heterocyclyls containing fused rings, one or more rings may be aryl or heteroaryl as defined below, insofar as the linkage points to the rest of the molecule are non-aromatic ring atoms. For the purposes of the present invention, the heterocyclyl is preferably a stable 4-11 member non-aromatic monoring containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur.

[0043] Co-drug In this specification, "co-drug," "co-carrier," or "interacting drug" can be used interchangeably and all refer to a single drug molecule that can be metabolized in the body to form two different drug molecules with different pharmacological effects. In this specification, a typical co-drug is the compound shown in formula I.

[0044] The aforementioned co-agents can be metabolized in the body to form various therapeutic agents. In the present invention, the preferred first therapeutic agent is berberine or an analog thereof, and the second therapeutic agent is a JAK family inhibitor.

[0045] First drug molecule In this specification, "first therapeutic agent" and "first drug molecule" both refer to the first drug molecule used in the co-agent of the present invention, and can be used interchangeably. The first drug molecule can lose any active functional group of its group to form a first drug group and link to the linking site of the co-agent molecule.

[0046] In the present invention, berberine or its analogues can be used as the first therapeutic agent of the co-agent. A preferred first drug molecule is represented by the following formulas II, III, or IV. [ka]

[0047] Second drug molecule In this specification, "second therapeutic agent" and "second drug molecule" both refer to the second drug molecule used in the co-agent of the present invention, and can be used interchangeably. The second drug molecule can lose one hydrogen atom of a group to form a second drug group and link to the linking site of the co-agent molecule.

[0048] A preferred second drug molecule is a JAK family inhibitor, which is clinically useful in treating intestinal diseases such as inflammatory bowel disease. The JAK family inhibitor may be a JAK inhibitor known in the art, or it may be a compound whose JAK inhibitory activity has not been verified.

[0049] The secondary drug group formed by exemplary JAK inhibitors is selected from the following group. [ka] [ka]

[0050] The compound of the present invention The compounds of the present invention are compounds represented by formula I, or their stereoisomers or racemates, or pharmaceutically acceptable salts thereof.

[0051] The compounds of the present invention may contain one or more chiral carbon atoms and thus can produce enantiomers, diastereomers, and other stereoisomers. Each chiral carbon atom may be defined as (R)- or (S)- based on stereochemistry. The present invention is intended to include all possible isomers, as well as their racemic and optically pure forms. The preparation of the compounds of the present invention can be done by selecting a racemic mixture, diastereomer, or enantiomer as a starting material or intermediate. Optically active isomers can be prepared using chiral synthtones or chiral reagents, or they can be resolved using prior art, for example, by methods such as crystallization and chiral chromatography.

[0052] Conventional techniques for preparing / separating individual isomers include chiral synthesis of suitable optically pure precursors or separation of racemates (or racemates of salts or derivatives) using, for example, chiral high-performance liquid chromatography. See, for example, Gerald Gubitz and Martin G. Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol. 243, 2004; AM Stalcup, Chiral Separations, Annu. Rev. Anal. Chem. 3:341-63, 2010; Fumiss et al. (eds.), VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup.TH ED., Longman Scientific and Technical Ltd., Essex, 1991, 809-816; Heller, Acc. Chem. Res. 1990, 23, 128.

[0053] The term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0054] A "pharmaceutically acceptable acid addition salt" refers to a salt of an inorganic or organic acid that can preserve the biological efficacy of a free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, and phosphate salts. Organic salts include, but are not limited to, formate, acetate, 2,2-dichloroacetate, trifluoroacetate, propionate, caproate, caprylate, caprine, undecylenate, glycolate, gluconate, lactate, sebacinate, adipine, glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate, cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, alginate, ascorbate, salicylate, 4-aminosalicylate, and naphthalenedisulfonate. These salts can be produced by methods known to those skilled in the art.

[0055] A "pharmaceutically acceptable base addition salt" is a salt with an inorganic or organic base that can preserve the biological efficacy of a free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, and aluminum salts. Preferred inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts, and magnesium salts. Salts derived from organic bases include, but are not limited to, primary amines, secondary amines, tertiary amines, substituted amines such as naturally substituted amines, cyclic amines, basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, and polyamine resins. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be produced by methods known to those skilled in the art.

[0056] Preparation method The following reaction scheme illustrates a method for preparing the compound represented by formula I, or its stereoisomers or racemates, or pharmaceutically acceptable salts thereof, where each group is as described above. In the reaction scheme below, the combination of substituents and / or variables in the general formula is acceptable only if such a combination results in a stable compound. It should also be understood that other general formulas may be prepared by technicians in the field of organic chemistry by the methods disclosed herein (using appropriately substituted starting materials and, if necessary, by modifying the synthetic parameters using methods well known to those skilled in the art) or by known methods.

[0057] In various embodiments and examples, the present invention provides tofacitinib glucuronide co-agents or pharmaceutically acceptable salts thereof, pharmaceutical compositions containing such compounds, methods for treating gastrointestinal inflammatory diseases using such compounds, and methods and intermediates for producing such compounds.

[0058] The compounds described herein may contain one or more chiral centers. In such cases, the description or naming of a particular stereoisomer means that the specified stereocenter has stereochemistry, and it should be understood here that, unless otherwise specified, a small number of other stereoisomers may be present, provided that their presence does not diminish the usefulness of the compound described or named.

[0059] Furthermore, as used herein, “compounds of the present invention” and “compounds of formula I” (or similar terms) are intended to include pharmaceutically acceptable salts unless otherwise specified.

[0060] use Because the co-pharmaceutical compounds of the present invention have excellent intestinal targeting effects, the compounds of the present invention, their various crystalline forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as major active ingredients are useful for the prevention and / or treatment of intestinal dysfunction, preferably gastrointestinal inflammatory diseases.

[0061] In this application, the term “pharmaceutical composition” means a formulation of the compound of the present invention with a medium generally accepted in the art for delivering a biologically active compound to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The pharmaceutical composition is intended to facilitate administration to a living organism, facilitate the absorption of the active ingredient, and further exert biological activity.

[0062] In this application, the term "pharmaceutically acceptable" means a substance (carrier or diluent) that does not affect the biological activity or properties of the compound of the present invention and is relatively non-toxic, that is, the substance can be applied to an organism without causing an undesirable biological reaction or interacting undesirably with any component contained in the composition.

[0063] In this application, the term “pharmaceutically acceptable excipient” includes, but is not limited to, any adjuvants, carriers, excipients, flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavorings, surfactants, wetting agents, dispersants, suspension aids, stabilizers, isotonic agents, solvents, or emulsifiers permitted by the relevant government authorities for use in humans or livestock.

[0064] In this application, the term "tumor" includes, but is not limited to, diseases such as glioma, sarcoma, melanoma, articular chondroma, cholangiomas, leukemia, gastrointestinal stromal malformation, histocellular lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, squamous cell carcinoma of the lung, adenocarcinoma of the lung, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, cervical cancer, ovarian cancer, intestinal cancer, nasopharyngeal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, and oral cancer.

[0065] In this application, the terms “preventive,” “prevention,” and “prevention” include diseases that reduce the likelihood of the onset or worsening of a disease or disorder.

[0066] In this application, the term "treatment" and other similar synonyms include the following meanings: (i) In particular, to prevent the occurrence of disease or disorder in mammals where the mammal is susceptible to the disease or disorder but has not been diagnosed with the disease or disorder. (ii) Inhibiting a disease or disorder, that is, inhibiting its onset, (iii) To alleviate a disease or disability, that is, to eliminate the state of the disease or disability or (iv) To alleviate symptoms caused by the disease or disorder.

[0067] In this application, the terms “effective dose,” “therapeutic effective dose,” or “pharmaceutical effective dose” mean an amount of at least one drug or compound sufficient to alleviate, to some extent, one or more symptoms of the disease or disorder being treated after administration. The result may be a reduction and / or alleviation of signs, symptoms, or etiology, or other desired changes in the biological system. For example, “effective dose” for treatment is the amount of a composition containing the compounds disclosed herein that is necessary to provide a clinically significant symptom-relieving effect. An effective dose suitable for any individual case can be determined using methods such as dose-escalation studies.

[0068] In this application, terms such as “administered,” “applied,” and “given” mean methods that enable the delivery of a compound or composition to a desired site for biological action. These methods include, but are not limited to, oral routes, duodenal routes, extra-gastrointestinal injections (including intravenous, subcutaneous, intraperitoneal, intramuscular, intra-arterial injections or infusions), topical administration, and rectal administration. The compounds and application techniques used in the methods described herein are known to those skilled in the art, for example, those discussed in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remingtons, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa. In preferred embodiments, the compounds and compositions described herein are applied orally.

[0069] In this application, the terms “combination of drugs,” “combination of drugs,” “combination,” “application of other treatments,” and “application of other therapeutic agents” refer to pharmaceutical treatments obtained by mixing or combining multiple active ingredients, and include combinations of immobilized and unimmobilized active ingredients. The term “immobilized combination” means administering at least one compound and at least one synergistic agent described herein to a patient simultaneously in the form of separate entities or separate dosage forms. The term “unimmobilized combination” means administering at least one compound and at least one synergistic formulation described herein to a patient simultaneously, in combination, or sequentially at variable intervals of time in the form of separate entities. These also apply to cocktail therapies, such as the application of three or more active ingredients.

[0070] Compared to conventional technology, the main advantages of the present invention are as follows: 1. The co-pharmaceutical compound of the present invention is not effectively absorbed by itself and can release two pharmacoactive components in the intestinal tract, thereby enriching the pharmacoactive components at the treatment site of the gastrointestinal tract and reducing systemic drug exposure. 2. The compounds of the present invention can effectively release JAK inhibitors (e.g., tofacitinib) and berberbine or its analogs in the intestines, thereby synergistically treating autoimmune inflammatory diseases of the gastrointestinal tract.

[0071] The present invention will be further described below with reference to specific examples. These examples are used solely for the purpose of illustrating the present invention and are not intended to limit its scope. Experimental methods in the following examples that do not specify concrete conditions generally follow normal conditions or conditions suggested by the manufacturer. Unless otherwise specified, percentages and quantities are calculated by weight.

[0072] In each embodiment, Analysis method I LCMS equipment: waters Acquity UPLC-MS, UV detector: Acquity UPLC Column: Acquity UPLC HSS T3 1.8uM, column temperature 40℃ Mobile phase: A: H2O (0.1% TFA), B: Acetonitrile, gradient elution

[0073] The preparation of intermediate A: (10-methoxy-9-((methyl(2-(methyl(((10-carbonyl-10-((5-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)-2-(((2S,3R,4S,5S,6S)-3,4,5-triacetoxy-6-(carbomethoxy<methoxycarbonyl>)tetrahydro-2H-pyran-2-yl)oxo)phenyl)amino)decyl)oxo)carbonyl)amino)ethyl)carbamoyl)oxo)-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation) was carried out according to the steps shown in the following formula. . [ka]

[0074] Intermediate A-1: ​​(2S,3R,4S,5S,6S)-2-(4-formyl-2-nitrophenoxy)-6-(carbomethoxy<methoxycarbonyl>)tetrahydro-2H-pyran-3,4,5-triyltriacetate [ka]

[0075] Under light-shielding conditions, the reactant (2R,3R,4S,5S,6S)-2-bromo-6-(carbomethoxy<methoxycarbonyl>)tetrahydro-2H-pyran-3,4,5-triyltriacetate (300 g, 755 mmol), the reactant 4-hydroxyl-3-nitrobenzaldehyde (214.6 g, 1284 mmol), and silver oxide (788 g, 3400 mmol) were added to 4 L of acetonitrile and stirred at 25-30°C for 5 hours. Monitoring by LC-MS confirmed that the starting materials had completely converted. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was diluted with ethyl acetate, filtered, and the filtrates were washed with saturated sodium bicarbonate solution and saturated brine, respectively. Liquid-liquid separation was performed, and the organic phase was dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure to obtain the title compound (295 g, 81%) as a yellow solid. MS (ESI): m / z = 506.1 [M+Na] + .

[0076] Intermediate A-2: (2S,3R,4S,5S,6S)-2-(4-(hydroxymethyl)-2-nitrophenoxy)-6-(carbomethoxy<methoxycarbonyl>)tetrahydro-2H-pyran-3,4,5-triyltriacetate [ka]

[0077] Intermediate A-1 (46.5 g, 96 mmol) and 19 g of silica gel were added to 450 ml of dichloromethane and 90 ml of isopropanol. The reaction was cooled to 0°C, and 5.5 g of sodium borohydride was slowly added. The reaction mixture was stirred at 0°C for 2 hours. Monitoring by LC-MS confirmed that the starting materials had completely converted. The reaction mixture was filtered, saturated ammonium chloride solution (200 ml) was added to the filtrate, and after liquid-liquid separation, the organic phase was washed twice with saturated brine (300 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was pulped with methyl t-butyl ether to obtain the title compound (340 g, 72.9%) as a white solid. MS (ESI): m / z = 508.1 [M+Na] + .

[0078] Intermediate A-3: (2S,3S,4S,5R,6S)-2-(carbomethoxy<methoxycarbonyl>)-6-(2-nitro-4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltriacetate [ka]

[0079] Intermediate A-2 (150 g, 310 mmol) and triethylamine (62.4 g, 620 mmol) were added to 1.5 L of dichloromethane. 4-nitrophenyl chloroformate (71.6 g, 350 mmol) was dissolved in 300 ml of dichloromethane and added dropwise to the reaction mixture at 0°C under nitrogen protection. After the addition was complete, the reaction mixture was stirred at 25°C for 6 hours. Monitoring by LC-MS confirmed that the starting materials had completely converted. t-butylmethyl (2-(methylamino)ethyl) carbamate (75.8 g, 400 mmol) was added dropwise to the reaction mixture from the previous step at 0°C. After the addition was complete, the reaction mixture was stirred at 25°C for 16 hours. Monitoring by LC-MS confirmed that the starting materials had completely converted. The reaction mixture was cooled to 0°C, 1 L of saturated sodium bicarbonate solution was added, and the organic phase was collected by liquid-liquid extraction. The organic phase was washed with saturated sodium bicarbonate solution (800 ml x 8), then with saturated brine (800 ml), dried over anhydrous sodium sulfate, and filtered. The organic phase was concentrated under reduced pressure to obtain the target compound (200 g, 92%). MS (ESI): m / z = 722.2 [M+Na] + .

[0080] Intermediate A-4: (2S,3R,4S,5S,6S)-2-(2-amino-4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenoxy)-6-(carbomethoxy<methoxycarbonyl>)tetrahydro-2H-pyran-3,4,5-triyltriacetate [ka]

[0081] Intermediate A-3 (200g, 285.8 mmol) was dissolved in 2 L methanol and 550 ml water, and iron powder (80g, 1429.1 mmol) and ammonium chloride (153g, 2858.1 mmol) were slowly added. The reaction mixture was stirred at 70°C under nitrogen protection for 5 hours. Monitoring by LC-MS confirmed that the starting materials had completely converted. The reaction mixture was filtered, and the filter cake was washed with 2 L ethyl acetate. The organic phase was concentrated under reduced pressure to obtain the crude product. 2 L ethyl acetate and 1.5 L water were added to the crude product, and the mixture was separated. The organic phase was washed three times with saturated brine (500 ml each time), and after separation, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane:methanol = 40:1) to obtain the target compound (100g, 52%) as a yellow oily substance. MS (ESI): m / z = 670.2 [M+H] +

[0082] 1 HNMR(400 MHz, DMSO-d6) δ 6.83 (d, J = 8.2 Hz, 1H), 6.66 (s, 1H), 6.50 (d, J = 8.2 Hz, 1H), 5.52 - 5.43 (m, 2H), 5.12 - 5.03 (m, 2H), 4.86 (s, 2H), 4.68 (d, J = 10.0 Hz, 3H), 3.64 (s, 3H), 3.31 - 3.25 (m, 3H), 2.79 (dd, J = 38.7, 13.2 Hz, 7H), 2.02 (d, J = 12.9 Hz, 9H), 1.36 (s, 9H).

[0083] Intermediate A-5: (2S,3R,4S,5S,6S)-2-(2-(10-hydroxyldecanoylamino)-4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenoxy)-6-(carbomethoxy<methoxycarbonyl>)tetrahydro-2H-pyran-3,4,5-triyltriacetate [ka]

[0084] Intermediate A-4 (100 g, 150 mmol) was dissolved in N,N-dimethylformamide (600 ml), and triethylamine (51.8 ml, 0.37 mmol) and the reactant 10-hydroxyldecanoic acid (39.3 g, 210 mmol) were added. Then, O-(7-azabenzotriazol-1-yl)-N,N,N'',N''-tetramethyluronium hexafluorophosphate (79.5 g, 210 mmol) was slowly added. After the addition was complete, the reaction mixture was stirred at 50°C for 16 hours under nitrogen protection. The reaction was stopped, and the reactant was diluted with 2 L of ethyl acetate. The diluted reaction mixture was washed with water (1.5 L * 8) and saline solution (1.5 L * 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane:methanol = 20:1) to obtain the title compound (47.0 g, 37%). MS (ESI): m / z = 862.2 [M+Na] + .

[0085] Intermediate A-6: (2S,3S,4S,5R,6S)-2-(carbomethoxy<methoxycarbonyl>)-6-(2-(10-((methyl(2-(methylamino)ethyl)carbamoyl)oxo)decanoylamino)-4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltriacetate [ka]

[0086] Intermediate A-5 (46.0 g, 57.7 mmol) and triethylamine (15.2 ml, 109.8 mmol) were dissolved in 400 ml of dichloromethane, and a solution of 4-nitrophenyl chloroformate (14.3 g, 71.2 mmol) in dichloromethane (60 ml) was added dropwise to the reaction mixture at 0°C under nitrogen protection. After the addition was complete, the reaction mixture was stirred at 25-30°C for 16 hours. Monitoring by LC-MS confirmed that the starting materials had been completely converted. Triethylamine (22.8 ml, 164.4 mmol) was added to the reaction mixture from the previous step. Then, N1,N2-dimethylethane-1,2-diamine (14.5 g, 164.4 mmol) was added dropwise to the reaction mixture from the previous step at 0°C under nitrogen protection. After the addition was complete, the mixture was stirred at 25-30°C for 4 hours. Upon monitoring by LC-MS, the reaction was completed, and the reaction solution was diluted with dichloromethane (800 ml). The diluted organic phase was washed with saturated sodium bicarbonate solution (600 ml x 3) and saline solution (700 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by normal-phase column chromatography (dichloromethane:methanol = 10:1) to obtain the title compound (36.0 g, 68%) as a yellow oily substance. MS (ESI): m / z = 954.5 [M+H] + .

[0087] Intermediate A-7: (2S,3R,4S,5S,6S)-2-(2-(10-(((2-((chlorocarbonyl)(methyl)amino)ethyl)(methyl)carbamoyl)oxo)decanoylamino)-4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenoxy)-6-(carbomethoxy<methoxycarbonyl>)tetrahydro-2H-pyran-3,4,5-triyltriacetate [ka]

[0088] Triphosgene (11.2 g, 37.7 mmol) was dissolved in 100 ml of dichloromethane in a three-necked flask, and intermediate A-6 (36.0 g, 37.7 mmol) was dissolved in 300 ml of dichloromethane. This solution was added dropwise to the triphosgene-dichloromethane solution at 0°C under nitrogen protection. After the addition was complete, the reaction mixture was stirred at room temperature for 10 minutes, and then triethylamine (15.7 ml, 113.2 mmol) was added dropwise to the reaction mixture at 0°C under nitrogen protection. After the addition was complete, the reaction mixture was stirred at 25-30°C for 3 hours. Monitoring by LC-MS confirmed that the starting materials had completely converted. The reaction mixture was cooled to 0°C, saturated sodium bicarbonate solution (300 ml) was added, and the mixture was separated. The organic phase was washed with saturated sodium bicarbonate solution (300 ml x 2) and saturated brine (200 ml). The compound was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the target compound (46.0 g of crude product), which was then used in the next step of the reaction without further purification. MS (ESI): m / z = 1038.3 [M+Na] + .

[0089] Intermediate A-8: 10-Methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation-9-alcoholic acid [ka]

[0090] Berberine hydrochloride (35.0 g, 94.3 mmol) was placed in a round-bottom flask and heated to 180°C under vacuum conditions using an oil pump. After 4 hours, it was cooled to room temperature. The crude product was pulped with ethanol, filtered, and dried to obtain the title compound (23.0 g, 73%) as a red solid. MS (ESI): m / z = 322.1 [M] + .

[0091] Intermediate A: 10-Methoxy-9-((methyl(2-(methyl(((10-carbonyl-10-((5-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)-2-(((2S,3R,4S,5S,6S)-3,4,5-triacetoxy-6-(carbomethoxy<methoxycarbonyl>)tetrahydro-2H-pyran-2-yl)oxo)phenyl)amino)decyl)oxo)carbonyl)amino)ethyl)carbamoyl)oxo)-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-cation [ka]

[0092] Intermediate A-8 (9.6 g, 30.0 mmol) was dissolved in 100 ml of pyridine in a three-necked flask, and intermediate A-7 (46.0 g, 45.0 mmol) was added dropwise to the three-necked flask in 300 ml of pyridine at 0°C under nitrogen protection. The reaction mixture was stirred at 25°C for 16 hours. Monitoring by LC-MS confirmed that the starting materials were completely converted. The reaction mixture was then concentrated under reduced pressure, and the crude product was purified by column chromatography (dichloromethane:methanol = 10:1) to obtain a black solid crude product. The crude product was purified by normal-phase column chromatography (dichloromethane:methanol = 3:2) to obtain the title compound (11.0 g, 22%). MS (ESI): m / z = 601.6 (M-100+H / 2) + .

[0093] 1 HNMR(400MHz, CDCl3) δ 11.24-10.55 (m, 1H), 8.45 (s, 2H), 7.83 (dd, J = 35.6, 27.7Hz, 3H), 7.42 (d, J = 20.4Hz, 1H), 7.21 - 6.74 (m, 3H), 6.27 (d, J = 4.1Hz, 0H), 6.08 (s, 2H), 5.78 (s, 0H), 5.57 - 5.23 (m, 6H), 5.06 (d, J = 11.4Hz, 2H), 4.23 - 3.95 (m, 5H), 3.78 (d, J = 18.4Hz, 3H), 3.54 - 2.73 (m, 20H), 2.34 (t, J = 23.9Hz, 2H), 2.09 (dt, J = 9.8, 4.0Hz, 8H), 1.88 - 0.86 (m, 27H).

[0094] Intermediate B: 2-Hydroxyl-5-((4-(4,7,10,10-tetramethyl-3,8-dioxy-2,9-dioxy-4,7-diazacyclo)phenyl)dinitro)benzoic acid [ka]

[0095] Intermediate B-1: 2-Hydroxyl-5-((4-(hydroxymethyl)phenyl)diazenyl)benzoic acid [ka]

[0096] A suspension of 4-aminobenzyl alcohol (2.0 g, 16.2 mmol) was treated with 3.4 ml of concentrated hydrochloric acid in 30 ml of water at 0°C, and then an aqueous solution of ice-cold NaNO2 (1.2 g, 17.0 mmol, 8 ml) was added. After stirring at 0°C for 1 hour, the reaction solution was added to an aqueous solution (25 ml) of sodium 2-hydroxybenzoate (2.72 g, 0.35 mmol) and potassium carbonate (3.2 g, 22.7 mmol). Throughout the dropwise addition process, an aqueous solution of sodium hydroxide was added dropwise to maintain the pH of the reaction solution at 13-14. The mixture was stirred at room temperature for 1 hour, the pH was adjusted to 4-5 with hydrochloric acid (2N), the product was precipitated, filtered and precipitated, washed with water (50 ml), and vacuum dried to obtain the title compound (4.0 g, 90%) as a red solid. MS (ESI): m / z = 272.8 [M+H] + .

[0097] Intermediate B: 2-Hydroxyl-5-((4-(4,7,10,10-tetramethyl-3,8-dioxy-2,9-dioxy-4,7-diazacyclo)phenyl)dinitro)benzoic acid [ka]

[0098] Intermediate B-1 (200 mg, 0.73 mmol) and diisopropylethylamine (114 mg, 0.88 mmol) were dissolved in 5 ml of dichloromethane, and bis(4-nitrophenyl) carbonate (268 mg, 0.88 mmol) was added. The reaction mixture was stirred at room temperature for 48 hours. t-butylmethyl (2-(methylamino)ethyl) carbamate (165 mg, 0.88 mmol) and diisopropylethylamine (114 mg, 0.88 mmol) were added dropwise to the reaction mixture from the previous step at 0°C. After the addition was complete, the reaction mixture was stirred at 25°C for 2 hours. Monitoring by LC-MS confirmed that the starting materials had completely converted. The reaction mixture was concentrated under reduced pressure and subjected to reverse-phase column chromatography to obtain the target compound (195 mg, 55%) as a red solid. MS (ESI): m / z = 508.9 [M+Na] + .

[0099] 1 H NMR (400 MHz, CDCl3) δ 8.29 (d, J = 2.4 Hz, 1H), 8.04 (dd, J1= 2.4 Hz, J2= 8.8 Hz, 1H), 7.82 (d, J = 8.4 Hz, 2H), 7.50 (d, J = 8.0 Hz, 2H), 7.11 (d, J = 9.2 Hz, 1H), 5.10 (s, 2H), 3.35-3.32 (m, 4H), 2.88-2.82 (m, 3H), 2.73-2.68 (m, 3H), 1.32 (s, 9H).

[0100] Intermediate C:9-(((2-(13-carboxyltridecanoylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0101] Intermediate C-1: 9-(((2-((t-butoxycarbonyl)amino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0102] t-butyl(2-(methylamino)ethyl)carbamate (15.0 g, 86.0 mmol) was dissolved in dichloromethane (200 ml), and triphosgene (25.6 g, 86.0 mmol) and pyridine (20.0 g, 258 mmol) were added sequentially in an ice bath. The mixture was stirred at room temperature (15 °C) for 1 hour. TLC detection confirmed that the reaction of the starting materials was complete. The reaction mixture was washed with water (200 ml), and the aqueous phase was extracted with dichloromethane (100 ml * 2). The organic phase was washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain an intermediate. The intermediate was dissolved in pyridine (20 ml), and intermediate A-8 (27.7 g, 86.0 mmol), which had been dissolved in pyridine (30 ml) in an ice bath, was added. The reaction was heated to room temperature (15 °C) and stirred for 16 hours. LC-MS detection confirmed that the reaction of the starting materials was complete. The reaction mixture was concentrated under reduced pressure and purified by normal-phase column chromatography (dichloromethane:methanol = 10:1) to obtain a yellow solid product (6.5 g, 14%). MS (ESI): m / z = 522.1[M] + .

[0103] Intermediate C: 9-(((2-aminoethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cationic hydrochloride [ka]

[0104] Intermediate C-1 (4.5 g, 8.6 mmol) was mixed with methanol hydrochloride solution (2 mol / L, 100 ml) and stirred overnight at room temperature (15°C). Detection by LC-MS confirmed the reaction was complete. The solution was concentrated under reduced pressure to obtain the title compound (3.6 g, 100%) as a brown solid. MS (ESI): m / z = 422.1 [M] + .

[0105] Intermediate D: 9-((4-hydroxylbenzyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0106] To a solution of intermediate A-8 (300 mg, 0.93 mmol) in acetonitrile (3 ml), 4-(chloromethyl)phenylacetate (257 mg, 1.4 mmol) and potassium carbonate (257 mg, 1.86 mmol) were added. The reaction was heated to 80°C and allowed to proceed for 16 hours. The reaction solution was diluted with dichloromethane (50 ml), filtered, and the filtered cake was washed with water (50 ml). The filtered cake was purified by normal-phase column chromatography (dichloromethane:methanol = 10:1) to obtain the title compound (97 mg, 24%) as a deep red solid. MS (ESI): m / z = 428.1 [M] + .

[0107] Preparation of Co-drug Compounds Example 1: 9-(((2-((((10-((2-(((2S,3R,4S,5S,6S)-6-carboxyl-3,4,5-trihydroxyltetrahydro-2H-pyran-2-yl)oxo)-5-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d] Pyrimidine-7-carboxamide (oxamide) ethyl)(methyl)carbamoyl) oxo)methyl)phenyl)amino)-10-carbonyldecyl) oxo)carbonyl)(methyl)amino) ethyl)(methyl)carbamoyl) oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0108] Examples 1-11: 4-Nitrophenyl 4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate [ka]

[0109] Tofacitinib (8.9 g, 28.6 mmol) was dissolved in dichloromethane solution (140 ml), and an aqueous solution of sodium hydroxide (3.4 g, 85.6 mmol) and tetrabutylammonium bromide (920 mg, 2.86 mmol) (48 ml) was added. A solution of p-nitrophenyl chloroformate (11.5 g, 57.1 mmol) in dichloromethane (48 ml) was added dropwise to the reaction mixture. After the addition was complete, the reaction mixture was stirred at room temperature for 4 hours. Monitoring by LC-MS confirmed that the starting materials had completely converted. The mixture was diluted with 500 ml of dichloromethane, washed with saturated ammonium chloride (200 ml), and insoluble matter was filtered off using diatomaceous earth. The organic phase was separated from the filtrate and then washed with 200 ml of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was pulped five times with dichloromethane and petroleum ether to obtain the title compound (16.3 g, 85%) as a yellow foamy solid. MS (ESI): m / z = 478.1 [M+H] + .

[0110] Example 1-10: 10-Methoxy-9-((methyl(2-(methyl(((10-((5-(((methyl(2-(methylamino)ethyl)carbamoyl)oxo)methyl)-2-(((2S,3R,4S,5S,6S)-3,4,5-triacetoxy-6-(carbomethoxy<methoxycarbonyl>)tetrahydro-2H-pyran-2-yl)oxo)phenyl)amino)-10-carbonyldecyl)oxo)carbonyl)amino)ethyl)carbamoyl)oxo)-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0111] To a solution of intermediate A (4.8 g, 3.7 mmol) in dichloromethane (40 ml), trifluoroacetic acid (10 ml) was added and the mixture was stirred at room temperature for 1 hour. The reaction was completed as monitored by LC-MS. The reaction mixture was concentrated under reduced pressure, dissolved in dichloromethane (30 ml), then concentrated under reduced pressure again to remove as much trifluoroacetic acid as possible, and dried by suction using an oil pump to obtain the title compound (4.43 g, 100%) as a yellow oily substance. MS (ESI): m / z = 601.4 [M / 2] + .

[0112] Examples 1-12: 9-(((2-((((10-((5-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)-2-(((2S,3R,4S,5S,6S)-3,4, 5-Triacetoxy-6-(carbomethoxy<methoxycarbonyl>)tetrahydro-2H-pyran-2-yl)oxo)phenyl)amino)-10-carbonyldecyl)oxo)carbonyl)(methyl)amino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0113] Solutions of Examples 1-10 (4.4 g, 3.7 mmol) in dichloromethane (100 ml) were cooled to 0°C, N,N-diisopropylethylamine (1.56 g, 12 mmol) was added, and then Examples 1-11 (1.76 g, 3.69 mmol) were added. The mixture was stirred at room temperature for 1 hour. The reaction was completed as monitored by LC-MS. The reaction solution was diluted with 300 ml of dichloromethane, washed sequentially with water and saturated saline solution, and the organic phase after washing was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was separated by normal-phase column chromatography (eluted with dichloromethane containing 7%-9% methanol) to obtain the title compound (2.83 g, 53%) as a yellow foamy solid. MS (ESI): m / z = 770.7 [M / 2] + .

[0114] Example 1: 9-(((2-((((10-((2-(((2S,3R,4S,5S,6S)-6-carboxyl-3,4,5-trihydroxyltetrahydro-2H-pyran-2-yl)oxo)-5-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d] Pyrimidine-7-carboxamide (oxamide) ethyl)(methyl)carbamoyl) oxo)methyl)phenyl)amino)-10-carbonyldecyl) oxo)carbonyl)(methyl)amino) ethyl)(methyl)carbamoyl) oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0115] A methanol (4 ml) solution of Examples 1-12 (163 mg, 0.105 mmol) was cooled to 0°C, and potassium carbonate aqueous solution (1 mmol / ml, 1 ml) was added. The mixture was stirred at 0°C for 2 hours. The reaction was completed as monitored by LC-MS. The pH of the reaction solution was adjusted to 5 with acetic acid, and then concentrated under reduced pressure. The crude product was separated by Prep-HPLC (gradient rinse with acetonitrile / water) to obtain the title compound (35.7 mg, 24%) as a yellow solid. MS (ESI): m / z = 1399.5 [M] + .

[0116] 1H NMR: (400 MHz, CD3OD) δ 10.06-9.59 (m, 1H), 8.71-8.61 (m, 1H),8.15-7.98 (m, 4H), 7.60-7.50 (m, 1H), 7.26-7.17 (m, 1H), 6.95-6.72 (m, 3H), 6.72-6.66 (m, 1H), 6.05 (s, 2H), 5.05-4.90 (m, 4H), 4.78-4.59 (m, 2H), 4.21-4.08 (m, 2H), 4.04 (s, 3H), 3.96-3.35 (m, 18H), 3.21-2.95 (m, 13H), 2.29-1.84 (m, 3H), 1.84-1.02 (m, 20H).

[0117] Each of the following compounds can be obtained by employing a method similar to that of Example 1, but by changing the corresponding raw materials. [Table 2]

[0118] Example 4A: 9-((5-((E)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation

[0119] Example 4B: 9-((5-((Z)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0120] Example 4-1: (E)-9-((2-hydroxyl-5-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)diazenyl)benzoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0121] Intermediate B (486 mg, 1.0 mmol), intermediate A-8 (322 mg, 1.0 mmol), and dicyclohexylcarbodiimide (247 mg, 1.2 mmol) were placed in a single-necked flask, and dichloromethane (10 ml) was added. The mixture was stirred at room temperature for 1 hour and monitored by LC-MS. Once the reaction was complete, the reaction mixture was filtered, concentrated under reduced pressure, and the crude product was separated by reverse-phase column chromatography to obtain the title compound (156 mg, 19.7%) as a brown oily substance. MS (ESI): m / z = 790.1 [M] + .

[0122] Example 4A: 9-((5-((E)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-cation and

[0123] Example 4B: 9-((5-((Z)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0124] Trifluoroacetic acid (0.4 ml) was added to a 2 ml solution of dichloromethane containing 156 mg of compound 4-1 (0.19 mmol), and the mixture was stirred at room temperature for 20 minutes. The reaction was completed as monitored by LC-MS. The reaction mixture was concentrated under reduced pressure, dried by suction using an oil pump, dissolved in a 2 ml solution of dichloromethane, cooled to 0°C, and N,N-diisopropylethylamine (101 mg, 0.78 mmol) was added. Compound 1-11 (94 mg, 0.19 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction was completed as monitored by LC-MS. The reaction mixture was concentrated under reduced pressure, and the crude product was separated by Prep-HPLC (gradient rinse with acetonitrile / water (containing 0.1% trifluoroacetic acid)) to obtain the title compound 4A (6.0 mg, 3.0%) as a yellow solid and 4B (6.0 mg, 3.0%) as a yellow solid. MS (ESI): m / z = 1028.4[M] + .

[0125] Example 5: 9-(((2-(5-((E)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0126] Example 5-1: (E)-9-(((2-(2-hydroxyl-5-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)diazenyl)benzoylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation

Chem.

[0127] Intermediate C (700 mg, 1.65 mmol), Intermediate B (782 mg, 1.65 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (477 mg, 2.48 mmol) were placed in a one-neck flask, N,N-dimethylformamide (5 ml) was added, and 4-dimethylaminopyridine (50 mg, 0.41 mmol) was added. The mixture was stirred at room temperature for 1 hour, and diisopropylethylamine (427 mg, 3.31 mmol) was added. The reaction solution was stirred at room temperature for 5 hours. When monitored by LCMS and the reaction was completed, 1N hydrochloric acid was added to quench it, the reaction solution was directly injected, separated by reverse-phase preparative chromatography, and the title compound (470 mg, 32%) was obtained as a yellow solid. MS (ESI): m / z = 890.3 [M] + .

[0128] Example 5-2: ((E)-9-(((2-(2-hydroxyl-5-((4-(((methyl(2-(methylamino)ethyl)carbamoyl)oxo)methyl)phenyl)diazenyl)benzoylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation

Chem.

[0129] A solution of Example 5-1 (470 mg, 0.53 mmol) in methanol (2 ml) was added with an ethyl acetate solution of hydrochloric acid (4 mol / L, 2 ml), and the mixture was stirred at room temperature for 1 hour. When monitored by LCMS, the reaction was completed. The reaction solution was concentrated under reduced pressure, dissolved in dichloromethane (10 ml), and then concentrated under reduced pressure again. This was repeated twice, and suction drying was carried out using an oil pump to obtain the title compound (390 mg, 93.5%) as a yellow solid. MS (ESI): m / z =790.2 [M+H] + .

[0130] Example 5: 9-(((2-(5-((E)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation

Chem.

[0131] A 5 ml solution of dichloromethane containing 390 mg of Example 5-2 (0.49 mmol) was cooled to 0°C, N,N-diisopropylethylamine (254 mg, 1.97 mmol) was added, and then Example 1-11 (235 mg, 0.49 mmol) was added. The mixture was stirred at room temperature for 1 hour. The reaction was completed as monitored by LC-MS. The reaction solution was diluted with 50 ml of dichloromethane, washed sequentially with 50 ml of water and 50 ml of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was subjected to normal-phase column chromatography (using 0.1% formic acid, dichloromethane, and methanol as eluents) to obtain the title compound (114 mg, 20%) as a yellow solid. MS (ESI): m / z = 1128.4 [M+H] + .

[0132] 1 H NMR: (400 MHz, CD3OD) δ 9.79-9.59 (m, 1H), 8.59-7.87 (m, 7H), 7.65-7.40 (m, 4H), 7.03-6.83 (m, 3H), 6.70-6.44 (m, 1H), 6.08 (s, 2H), 5.32-4.94 (m, 3H), 3.91-3.72 (m, 6H), 3.62-3.43 (m, 9H), 3.35 (s, 3H), 3.22-2.66 (m, 13H), 2.41-2.26 (m, 1H), 1.84-1.52 (m, 2H), 1.36-1.26 (m, 4H), 1.03-0.85 (m, 3H).

[0133] Example 6: 9-(((2-(6-(5-((E)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoylamino)caproylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0134] Example 6-1: Methyl(E)-6-(2-hydroxyl-5-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)diazenyl)benzoylamino)hexanoate [ka]

[0135] Intermediate B (1900 mg, 3.9 mmol), methyl 6-aminocaproate hydrochloride (849 mg, 4.6 mmol), and diisopropylethylamine (2017 mg, 15.6 mmol) were dissolved in N,N-dimethylformamide (19 ml), and O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate (2228 mg, 5.86 mmol) was added. The mixture was stirred at 30 °C for 3 hours. When the reaction was completed as monitored by LCMS, the reaction solution was diluted with ethyl acetate (150 ml), washed 4 times with water (100 ml), washed once with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by normal-phase column chromatography (petroleum ether:ethyl acetate = 2:1) to obtain the title compound (870 mg, 36.3%) as a yellow solid. MS (ESI): m / z = 636.2 [M+Na] + .

[0136] Example 6-2: (E)-6-(2-hydroxyl-5-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)diazenyl)benzoylamino)caproic acid

Chemical formula

[0137] Example 6-1 (800 mg, 1.3 mmol) was dissolved in methanol (5 ml) and water (2 ml), and lithium hydroxide monohydrate (247 mg, 6.5 mmol) was added. The reaction solution was stirred at 65 °C for 1 hour. When the reaction was completed as monitored by LCMS, it was cooled to room temperature, neutralized to a pH value of 4 - 5 with dilute hydrochloric acid, extracted with 100 ml of ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the title compound (784 mg, 100%) was obtained as a yellow solid. MS (ESI): m / z = 622.2 [M+Na] + .

[0138] Example 6-3: (E)-9-(((2-(6-(2-hydroxyl-5-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)diazenyl)benzoylamino)caproylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation

Chemical formula

[0139] To a mixture of Example 6-2 (820 mg, 1.37 mmol), intermediate C (693 mg, 1.64 mmol), and DMF (1 ml), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (780 mg, 2.05 mmol) and diisopropylethylamine (706 mg, 5.47 mmol) were added sequentially. After the addition was complete, the mixture was stirred at room temperature for 0.5 hours. When the reaction was completed as monitored by LC-MS, the reaction mixture was diluted with dichloromethane (100 ml), washed with water (50 ml * 4) and saline solution (50 ml * 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (methanol:dichloromethane = 1:12, (containing 0.1% formic acid)) to obtain the title compound (525 mg, 38%) as a yellow solid. MS (ESI): m / z = 1003.3 [M] + .

[0140] Example 6: 9-(((2-(6-(5-((E)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoylamino)caproylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0141] To a solution of Example 6-3 (525 mg, 0.52 mmol) in dichloromethane (2 ml), trifluoroacetic acid (0.4 ml) was added and the mixture was stirred at room temperature for 1 hour. The reaction was completed as monitored by LC-MS. The reaction mixture was concentrated under reduced pressure, dried by suction using an oil pump, and then dissolved in dichloromethane (2 ml). The solution was cooled to 0°C, N,N-diisopropylethylamine (270 mg, 2.09 mmol) was added, and Example 1-11 (249 mg, 0.52 mmol) was added. The mixture was stirred at room temperature for 0.5 hours. The reaction was completed as monitored by LC-MS. The reaction mixture was concentrated under reduced pressure, and the crude product was separated by Prep-HPLC (gradient rinse with acetonitrile / water (containing 0.1% trifluoroacetic acid)) to obtain the title compound (135 mg, 21%) as a yellow solid. MS (ESI): m / z = 1241.8[M] + .

[0142] 1 H NMR: (400 MHz, CD3OD) δ 9.89-9.59 (m, 1H), 8.59-8.47 (m, 2H), 8.37-8.31 (m, 1H), 8.12-7.93 (m, 2H), 7.70-7.61 (m, 2H), 7.51-7.43 (m, 2H), 6.75-6.73 (m, 1H), 6.05 (s, 2H), 5.17-4.90 (m, 4H), 3.98 (s, 3H), 3.79-3.25 (m, 14H), 3.23-3.06 (m, 9H), 2.32-2.27 (m, 3H), 1.72-1.21 (m, 11H), 0.99-0.87 (m, 4H).

[0143] Example 7: 9-(((2-(14-((4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)amino)-14-carbonyltetradecanoylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0144] Example 7-1: 4-(((t-butyldimethylsilyl)oxo)methyl)aniline [ka] (4-aminophenyl)methanol (5.0 g, 40.6 mmol) and imidazole (3.04 g, 44.66 mmol) were dissolved in dichloromethane (70 ml), to which t-butyldimethylchlorosilane (6.12 g, 40.6 mmol) was added, and the mixture was reacted at room temperature for 1 hour. Ethyl acetate (200 ml) was added to the reaction mixture, the organic phase was washed with water (400 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by normal-phase column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain the title compound (9.3 g, 95%) as a pale yellow liquid. MS (ESI): m / z = 238.1 [M+H] + .

[0145] Example 7-2: 14-((4-(((t-butyldimethylsilyl)oxo)methyl)phenyl)amino)-14-carbonylmyristic acid [ka] To a solution of Example 7-1 (4280 mg, 18.06 mmol) and tetrasebacic acid (6989 mg, 27.09 mmol) in dichloromethane (43 ml), N,N-diisopropylethylamine (4659 mg, 36.12 mmol) was added, and the mixture was cooled to 0°C and stirred for 20 minutes. O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (10.3 g, 27.09 mmol) was added to the reaction mixture. The reaction was heated to room temperature and stirred for 16 hours. The reaction mixture was diluted with dichloromethane (20 ml) and washed with water (40 ml). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was separated and purified by normal-phase column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain the title compound (6.7 g, 79%) as a pale yellow solid. MS (ESI): m / z = 500.2 [M+Na] + .

[0146] Example 7-3: 9-(((2-(14-((4-(((t-butyldimethylsilyl)oxo)methyl)phenyl)amino)-14-carbonyltetradecanoylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0147] A 20 ml solution of dichloromethane containing Example 7-2 (1800 mg, 3.77 mmol) and intermediate C (1592 mg, 3.77 mmol) was placed in a single-necked flask. N,N-diisopropylethylamine (1947 mg, 15.09 mmol) was added at 0°C, and the mixture was stirred for 20 minutes. Then, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2870 mg, 7.55 mmol) was added. The reaction mixture was heated to room temperature and stirred for 40 minutes. The reaction mixture was washed with water (40 ml), the organic phase was dried over anhydrous sodium sulfate, and the mixture was concentrated under reduced pressure. The residue was separated and purified by normal-phase column chromatography (ethyl acetate:petroleum ether = 1:1) to obtain the target compound (2200 mg, 66.2%) as a white solid. MS (ESI): m / z = 881.4 [M] + .

[0148] Example 7-4: 9-(((2-(14-((4-(hydroxymethyl)phenyl)amino)-14-carbonyltetradecanoylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0149] To 30 ml of tetrahydrofuran (2100 mg, 2.38 mmol) from Example 7-3, pyridine hydrogen fluoride (753 mg, 9.53 mmol) was added. The reaction was stirred at room temperature for 16 hours and monitored by LC-MS. Once the reaction was complete, it was diluted with dichloromethane (30 ml), washed with water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the target compound (1360 mg, 74.3%) as a white solid. MS (ESI): m / z = 767.3 [M] + .

[0150] Example 7-5: 10-Methoxy-9-((methyl(2-(14-carbonyl-14-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)amino)tetradecanoylamino)ethyl)carbamoyl)oxo)-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0151] Triethylamine (267 mg, 2.64 mmol) was slowly added at 0°C to 10 ml of dichloromethane (675 mg, 0.88 mmol) of Example 7-4. A solution of 4-nitrobenzoyl chloride (265 mg, 1.32 mmol) in dichloromethane (1 ml) was slowly added dropwise, and the reaction was stirred at room temperature for 1 hour. The reaction was then cooled to 0°C, and a solution of t-butylmethyl (2-(methylamino)ethyl) carbamate (249 mg, 1.32 mmol) in dichloromethane (1.32 ml) was slowly added dropwise, and the reaction was stirred at room temperature for 1 hour. The reaction mixture was washed with water (20 ml x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column (ethyl acetate:petroleum ether = 1:1) to obtain the target compound (240 mg, 28%) as an orange-yellow oily substance. MS (ESI): m / z = 981.5 [M] + .

[0152] Example 7: 9-(((2-(14-((4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)amino)-14-carbonyltetradecanoylamino)ethyl)(methyl)carbamoyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0153] A solution of dichloromethane (210 mg, 0.214 mmol) from Example 7-5 was placed in a 2 ml single-necked flask, trifluoroacetic acid (0.4 ml) was added, and the reaction mixture was stirred at room temperature for 15 minutes. When the reaction was completed, monitored by LC-MS, the reaction mixture was concentrated under reduced pressure, and dichloromethane (1 ml), N,N-diisopropylethylamine (0.1 ml, 0.64 mmol), and Example 1-11 (102 mg, 0.21 mmol) were added and the mixture was reacted at room temperature for 15 minutes. The reaction mixture was concentrated under reduced pressure, N,N-dimethylformamide (2 ml) was added, and the mixture was directly injected. Separation was performed by Prep-HPLC (gradient rinse with acetonitrile / water (containing 0.1% trifluoroacetic acid)) to obtain the target compound (15.2 mg, 5.8%) as a yellow solid. MS (ESI): m / z = 1219.7 [M] + .

[0154] 11H NMR (400 MHz, CD3OD) δ 10.039 - 9.792 (m, 1H), 8.76 - 8.73 (m, 1H), 8.53 (s, 1H), 8.15 (m, 3H), 7.64 - 7.62 (m, 1H), 7.51 - 7.50 (m, 1H), 7.46 - 7.44 (m, 1H), 7.29 - 7.27 (m, 1H), 6.93 - 6.91 (m, 1H), 6.78 - 6.66 (m, 2H), 6.082 (s, 2H), 5.00 - 4.99 (m, 5H), 4.06 (s, 3H), 3.96 - 3.87 (m, 4H), 3.68 (s, 3H), 3.57 - 3.55 (m, 2H), 3.48 - 3.44 (m, 4H), 3.23 - 3.21 (m, 3H), 3.08 (s, 2H), 2.42 - 2.40 (m, 1H), 2.33 - 2.29 (t, J = 8.0 Hz, 3H), 2.25 - 2.17 (m, 3H), 1.71 - 1.54 (m, 7H), 1.28 - 1.20 (m, 21H), 1.09 - 0.99 (m, 5H).

[0155] Example 8: 9-((5-(5-((E)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoylamino)valeryl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation

Chem.

[0156] Example 8-1: Methyl(E)-5-(2-hydroxyl-5-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)diazenyl)benzoylamino)valerate

Chem.

[0157] A solution of intermediate B (3.0 g, 6.17 mmol), 1-hydroxylbenzotriazole (1.001 g, 7.41 mmol), and dicyclohexylcarbodiimide (1.783 g, 8.64 mmol) in dichloromethane (20 ml) was stirred at 0°C for 20 minutes. Methyl 5-aminovalerate hydrochloride (1.212 g, 7.41 mmol) and N,N-diisopropylethylamine (1752 mg, 13.6 mmol) were added, and the reaction was allowed to proceed at room temperature for 2 hours. When the reaction was complete, as monitored by LC-MS, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by normal-phase column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain the title compound (3.34 g, 91%) as a red solid. MS (ESI): m / z = 622.2 [M+Na] + .

[0158] Example 8-2: (E)-5-(2-hydroxyl-5-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)diazenyl)benzoylamino)valeric acid [ka]

[0159] To a solution of Example 8-1 (3300 mg, 5.51 mmol) in methanol (30 ml) and water (15 ml), lithium hydroxide monohydrate (1157 mg, 27.5 mmol) was added and the mixture was heated under reflux at 65°C for 1 hour. After cooling, the pH of the reaction solution was adjusted to 5 with dilute hydrochloric acid. The mixture was extracted three times with ethyl acetate (100 ml), the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound (3000 mg, 93.1%) as an orange solid. MS (ESI): m / z = 608.2 [M+Na] + .

[0160] Example 8-3: (E)-9-((5-(2-hydroxyl-5-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)diazenyl)benzoylamino)valeryl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0161] A 20 ml solution of dichloromethane (1500 mg, 2.56 mmol) from Example 8-2 was placed in a single-necked flask, and dicyclohexylcarbodiimide (792 mg, 3.84 mmol) and intermediate A-8 (908 mg, 2.82 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. When the reaction was complete, as monitored by LC-MS, the reaction mixture was washed three times with water (60 ml), the organic phase was dried over anhydrous sodium sulfate, and the mixture was concentrated under reduced pressure. The residue was separated and purified by normal-phase column chromatography (dichloromethane:methanol = 10:1) to obtain the title compound (367 mg, 16.1%) as an orange oily substance. MS (ESI): m / z = 889.4 [M] + .

[0162] Example 8: 9-((5-(5-((E)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoylamino)valeryl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0163] Trifluoroacetic acid (0.5 ml) was added to dichloromethane (2.5 ml) of Example 8-3 (300 mg, 0.34 mmol). The reaction was stirred at room temperature for 15 minutes, the solution was concentrated under reduced pressure, and N,N-dimethylformamide (2 ml), N,N-diisopropylethylamine (95.8 mg, 0.74 mmol), and Example 1-11 (193 mg, 0.4 mmol) were added. The reaction was allowed to proceed at room temperature for 15 minutes. After detecting the end of the reaction by LC-MS, 1N hydrochloric acid was added to quench the reaction mixture, and the reaction solution was injected as is. Separation was performed by Prep-HPLC (gradient rinse with acetonitrile / water (containing 0.1% trifluoroacetic acid)) to obtain the target compound (62.0 mg, 16%) as a yellow solid. MS (ESI): m / z = 1127.5 [M] + .

[0164] 1H NMR (400 MHz, CD3OD) δ 9.63-9.60 (m, 1H), 8.70-8.62 (m, 1H), 8.39-8.35 (m, 1H), 8.11-8.02 (m, 3H), 7.91-7.89 (m, 1H), 7.68-7.66 (m, 2H), 7.59-7.56 (m, 2H), 7.46-7.40 (m, 2H), 7.01-6.85 (m, 3H), 6.09 (s, 2H), 5.18-5.16 (m, 2H), 4.03-4.02 (m, 3H), 3.92-3.91 (m, 3H), 3.81-3.73 (m, 3H), 3.65-3.59 (m, 2H), 3.56-3.52 (m, 4H), 3.46-3.39 (m, 2H), 3.24-3.18 (m, 5H), 3.17-3.16 (m, 3H), 2.94-2.91 (m, 4H), 2.36-2.28 (m, 1H), 1.99-1.94 (m, 2H), 1.91-1.86 (m, 2H), 1.68-1.60 (m, 5H), 0.94-0.92 (m, 2H).

[0165] Each of the following compounds can be obtained by employing a method similar to that of Example 8, but by changing the corresponding raw materials. [Table 3]

[0166] Example 10: 9-((3-(2-((E)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)phenyl)propionyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0167] Example 10-1: 1-Hydroxyl-3,4-dihydroquinoline-2(1H)-one [ka] Hydrogen peroxide (35%, 22 ml) was added dropwise to a methanol solution (200 ml) of 1,2,3,4-tetrahydroquinoline (10.0 g, 75 mmol) and sodium tungstate dihydrate (1.9 g, 3.7 mmol). After the addition was complete, the mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure, the residue was dissolved in water (200 ml), and extracted with dichloromethane (100 ml x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was pulped with dichloromethane / methanol (1:1, 100 ml) and filtered. The filtered cake was dried to obtain the title compound (8.8 g, 72%) as a yellowish-brown solid. MS (ESI): m / z = 164.1 [M+H] + .

[0168] 1 H NMR (CDCl3, 400 MHz) δ 9.03 (br, 1H), 7.33 (d, J = 8.0 Hz, 1H), 7.28 (t, J = 8.0 Hz, 1H), 7.15 (d, J = 8.8 Hz, 1H), 7.05 (t, J = 7.6 Hz, 1H), 2.93 (t, J = 7.6 Hz, 2H), 2.76 (t, J = 8.0 Hz, 2H).

[0169] Example 10-2: 3-(2-nitrosophenyl)propionic acid [ka] While cooling in an ice bath, sodium periodate (23.1 g, 107 mmol) was added to the solution of tetrahydrofuran (120 ml) and water (30 ml) of Example 10-1 (8.8 g, 53 mmol). The mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to remove the organic solvent, the residue was diluted with water (100 ml), the pH was adjusted to weakly acidic with hydrochloric acid (2 M), and the mixture was extracted with ethyl acetate (450 ml). After combining the organic phases, the mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting crude product was pulped with ethyl acetate (100 ml) to obtain the title compound (4.5 g, 46%) as a yellow solid. MS (ESI): m / z = 171.2 [M+H] + .

[0170] Example 10-3: (E)-3-(2-((4-(hydroxymethyl)phenyl)diazenyl)phenyl)propionic acid [ka] Example 10-2 (4.5 g, 25.1 mmol) and tetraaminobenzyl alcohol (3.1 g, 25.1 mmol) were dissolved in dichloromethane (150 ml), acetic acid (15 ml) was added, and the mixture was stirred at room temperature for 48 hours under nitrogen protection. The reaction was completed when monitored by LC-MS of the starting materials. The reaction solution was diluted with dichloromethane (100 ml), washed sequentially with water (100 ml x 2) and saturated brine (100 ml), the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by normal-phase column chromatography (dichloromethane:methanol = 94:6). The mixture was pulped with ethyl acetate (30 ml) to obtain the title compound (4.0 g, 56%) as a red solid. MS (ESI): m / z = 285.1 [M+H] + .

[0171] Example 10-4: (E)-3-(2-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)diazenyl)phenyl)propionic acid [ka]

[0172] Example 10-3 (3.7 g, 13.0 mmol) and bis(p-nitrophenyl) carbonate (4.752 g, 15.6 mmol) were mixed in dichloromethane (50 ml), and diisopropylethylamine (3.361 g, 26.0 mmol) was added dropwise while cooling in an ice bath. The reaction mixture was stirred at room temperature for 30 minutes, and then t-butylmethyl (2-(methylamino)ethyl) carbamate (3.184 g, 16.9 mmol) was added dropwise to the reaction mixture at 0°C. After the addition was complete, the reaction mixture was allowed to react at room temperature for 1 hour, then the temperature was raised to 40°C and stirred for 2 hours. Monitoring by LCMS confirmed that the starting materials had completely converted. The reaction mixture was diluted with dichloromethane (200 ml), washed with water (100 ml), the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by normal-phase column chromatography (dichloromethane:methanol = 10:1) to obtain the target compound (1.1 g, 17%) as a yellowed solid. MS (ESI): m / z = 521.1 [M+Na] + .

[0173] Example 10-5: (E)-10-Methoxy-9-((3-(2-((4-(4,7,10,10-tetramethyl-3,8-dicarbonyl-2,9-dioxa-4,7-diazaundecyl)phenyl)diazenyl)phenyl)propionyl)oxo)-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0174] In Example 10-4, (650 mg, 1.3 mmol) and pyridine (412 mg, 5.2 mmol) were mixed with dichloromethane (30 ml), and oxalyl chloride (486 mg, 1 mmol) was added dropwise at room temperature. The reaction mixture was stirred at room temperature for 30 minutes and monitored by LC-MS, where it was confirmed that the starting materials had converted. The reaction mixture was concentrated under reduced pressure, dried by suction using an oil pump, and then dissolved in acetonitrile (5 ml). This was then added dropwise to a solution of intermediate A-8 (420 mg, 1.3 mmol) and pyridine (412 mg, 5.2 mmol) in acetonitrile (30 ml). The reaction mixture was stirred at room temperature for 20 minutes and monitored by LC-MS, where it was confirmed that the starting materials had completely converted. The reaction mixture was concentrated under reduced pressure, diluted with dichloromethane (100 ml), washed with water (100 ml) and dilute hydrochloric acid (1 N, 100 ml), the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by normal-phase column chromatography (dichloromethane:methanol = 92:8, containing 0.1% trifluoroacetic acid) to obtain the target compound (217 mg, 21%) as a yellowish oily substance. MS (ESI): m / z = 803.2 [M+H] + .

[0175] Example 10: 9-((3-(2-((E)-(4-((((2-(4-(((3R,4R)-1-(2-cyanoacetyl)-4-methylpiperidine-3-yl)(methyl)amino)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)phenyl)propionyl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0176] In a 2 ml solution of dichloromethane (217 mg, 0.27 mmol) from Example 10-5, trifluoroacetic acid (0.4 ml) was added and the mixture was stirred at room temperature for 1 hour. The reaction was completed as monitored by LC-MS. The reaction mixture was concentrated under reduced pressure, dried by suction using an oil pump, dissolved in a 1.5 ml solution of N,N-dimethylformamide, cooled to 0°C, and N,N-diisopropylethylamine (139 mg, 1.1 mmol) was added. Then, Example 1-11 (258 mg, 0.54 mmol) was added and the mixture was stirred at room temperature for 0.5 hours. After quenching the reaction mixture with 1N hydrochloric acid, the mixture was injected as is and separated by Prep-HPLC (gradient rinse with acetonitrile / water (containing 0.1% trifluoroacetic acid)) to obtain the title compound (63.0 mg, 22.4%) as a yellow solid. MS (ESI): m / z = 1140.4 [M] + .

[0177] 1 H NMR (400 MHz, CD3OD) δ 9.60-9.47 (m, 1H), 8.72-8.67 (m, 1H), 8.19-8.07 (m, 3H), 7.87-7.74 (m, 2H), 7.63-7.39 (m, 6H), 6.99-6.51 (m, 4H), 6.09 (s, 2H), 5.21-5.05 (m, 1H), 4.97-4.86 (m, 3H), 4.00-3.71 (m, 8H), 3.66-3.37 (m, 8H), 3.24-3.16 (m, 9H), 3.05-2.81 (m, 6H), 2.40-2.26 (m, 1H), 1.90-1.77 (m, 1H).

[0178] Example 11: (E)-9-((5-(5-((4-((((2-(4-(1-(3-(cyanomethyl)-1-(ethylsulfonyl)azetidine-3-yl)-1H-pyrazole-4-yl)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoylamino)valeryl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0179] Example 8-3-A: (E)-9-((5-(2-hydroxyl-5-((4-(((methyl(2-(methylamino)ethyl)carbamoyl)oxo)methyl)phenyl)diazenyl)benzoylamino)valeryl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cationic hydrochloride [ka]

[0180] To a 35 ml solution of ethyl acetate (7.7 g, 7.13 mmol) of Example 8-3, ethyl acetate hydrochloride solution (4 N, 70 ml) was added at 0°C, and the mixture was reacted at 25°C for 1 hour. When the reaction was complete, monitored by LC-MS, the reaction solution was filtered, the filter cake was rinsed with ethyl acetate, and the title compound (6.8 g, 110%) was obtained as a red solid. MS (ESI): m / z = 789.3 [M] + .

[0181] Example 1-A: 4-Nitrophenyl 4-(1-(3-(cyanomethyl)-1-(ethylsulfonyl)azetidine-3-yl)-1H-pyrazole-4-yl)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate [ka]

[0182] p-nitrophenyl chloroformate (108 mg, 0.54 mmol) was added at room temperature to a mixture of baricitinib (100 mg, 0.27 mmol), triethylamine (109 mg, 1.08 mmol), and dichloromethane (5 ml). The reaction mixture was stirred at room temperature for 1.5 hours. Monitoring by LC-MS confirmed that the starting materials had completely converted. The reaction mixture was diluted with dichloromethane (50 ml), washed with water (50 ml) and saline solution (50 ml), respectively, and the organic phase was dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure to obtain the crude product. The crude product was pulped with dichloromethane (5 ml), filtered, and the filtered cake was dried to obtain the title compound (100 mg, 85%) as a yellow solid. MS (ESI): m / z = 537.0 [M+H] + .

[0183] Example 11: (E)-9-((5-(5-((4-((((2-(4-(1-(3-(cyanomethyl)-1-(ethylsulfonyl)azetidine-3-yl)-1H-pyrazole-4-yl)-N-methyl-7H-pyrrolo[2,3-d]pyrimidine-7-carboxamide<oxamide>)ethyl)(methyl)carbamoyl)oxo)methyl)phenyl)diazenyl)-2-hydroxylbenzoylamino)valeryl)oxo)-10-methoxy-5,6-dihydro-[1,3]dioxazolo[4,5-g]isoquinolino[3,2-a]isoquinoline-7-cation [ka]

[0184] To a mixture of Example 8-3-A (118 mg, 0.15 mmol), N-methylmorpholine (19 mg, 0.18 mmol), and N,N-dimethylformamide (2 ml), Example 1-A (100 mg, 0.18 mmol) was added. The mixture was stirred at room temperature for 2 hours. Monitoring by LC-MS confirmed that the starting materials had completely converted. The reaction mixture was quenched with hydrochloric acid (1 N). The reaction mixture was injected as is and separated by Prep-HPLC (gradient rinse with acetonitrile / water (containing 0.1% trifluoroacetic acid)) to obtain the title compound (93 mg, 42%) as a yellow solid. MS (ESI): m / z = 1186.4 [M] + .

[0185] 1 H NMR (400 MHz, CD3CN) δ 9.37(s, 1H), 8.98-8.55 (m, 4H), 8.18-8.02 (m, 4H), 7.66-7.33 (m, 6H), 7.04-6.88 (m, 3H), 6.12 (s, 2H), 5.29-5.09 (m, 1H), 4.77-4.55 (m, 5H), 4.28-4.22 (m, 3H), 4.02 (s, 3H), 3.77-3.46 (m, 9H), 3.12-2.91 (m, 14H), 1.34-1.31 (m, 5H).

[0186] The following compounds can be obtained by employing a method similar to that of Example 11, but by changing the corresponding starting materials. The reaction solvent for preparing Example 1-A may be dichloromethane, N,N-dimethylformamide, etc., and the base may be triethylamine, 2,6-dimethylpyridine, etc. The reaction solvent for preparing Example 11 may be N,N-dimethylformamide, N,N-dimethylacetamide, etc. [Table 4-1] [Table 4-2] [Table 4-3]

[0187] Biological test 1: Ex vivo assay of the duodenal or colon contents of mice containing the co-drug. Male C57 BL / 6 mice (6-8 weeks old) were euthanized with carbon dioxide and then dissected. The duodenum and colon were removed and placed in 1.5 ml centrifuge tubes, and PBS solution was added simultaneously. The intestinal portion was cut longitudinally, shaken to release the intestinal contents, and then mixed upside down. Dimethyl sulfoxide solutions of the compounds of the example were prepared separately, and 20 μL of each dimethyl sulfoxide solution was taken and added to 1 ml of duodenal or colon contents in PBS solution. The mixture was repeatedly mixed upside down and placed in a 37°C water bath. The above solutions were taken at 0, 1, 4, 16, and 20 hours, acetonitrile was added, the solutions were vortexed, and centrifuged for 10 minutes. The supernatant was taken and 10 μL of the internal standard compound (intermediate D) was added. The amounts (ng) of the compounds of the example, berberbine, and tofacitinib were detected using a liquid chromatography-mass spectrometer and quantified using a calibration curve. The results are shown in Table 1. As a result, it was shown that the co-pharmaceutical compound of the present invention can be released in the intestinal tract after administration.

[0188] [Table 5] ND not detected

[0189] Biological Experiment 2: Pharmacokinetic Experiment of Co-Drug Compounds in Mice The test compound was orally administered to CD-1 mice (PO, 15 mg / kg), and blood samples and tissue samples from various parts of the digestive tract were collected at different time points. The concentrations of the co-agent compound in the mouse plasma, as well as the released tofacitinib and berberbine, were measured by LC-MS / MS. The animals were approximately 6-8 weeks old at the start of the administration experiment. Blood and tissue samples were collected at 0.5, 1, 2, 4, 8, and 24 hours after administration. Analytical methods and detection methods for biological samples were established. The results are shown in Figures 1-5. The results show that the co-drug compound in the example was able to release tofacitinib and berberbine in the mouse intestine, and moreover, the co-drug compound, tofacitinib, and berberbine were all mainly restricted to the intestinal tissue, resulting in extremely low drug exposure in plasma.

[0190] Biological Experiment 3: Efficacy Experiment of Co-drug Compounds in an Oxazolone-Induced Mouse Colitis Model A model of oxazolone-induced colitis was created using C57 BL / 6 mice, following the method of Heller et al. On day 1, the skin of the dorsal neck of the mice was shaved (2 cm × 2 cm), and 150 ul of 3% oxazolone solution (dissolved in a 4:1 mixture of acetone and olive oil) was applied to sensitize the mice. Six days after sensitization, the mice were randomly divided into groups. Subsequently, the compound from the corresponding example (120 mg / kg) was administered intragastricly, while the blank control group and model group were administered the solvent. The intragastric administration rate was 10 ml / kg body weight. On day 2, 50 ul of 1.2% oxazolone solution was administered enema, and pure water was injected into the blank control group. Intragastric administration was continued for four days, and the Disease Activity Index (DAI) was recorded daily. As shown in Figure 6, the group administered the co-drug compound from the example showed a significant improvement in the Disease Activity Index compared to the model group.

[0191] All documents referenced in this invention are cited in this application to the same extent that each document is cited individually. Furthermore, those skilled in the art will understand, after reading the above, that various changes or modifications can be made to the invention, and that these equivalent forms are also included within the scope defined by the claims attached to this application.

Claims

1. A co-drug compound represented by formula I, formed by coupling a first drug molecule, a second drug molecule, and a linker precursor, 【Chemistry 1】 During the ceremony, D １ This is the first drug group, D ２ This is the second drug group, Furthermore, the linker is selected from the following groups (A), (B), or (c), and in each formula, J １ It is linked to the first drug group, J ２ It is linked to the second drug group, (A) Group is -L ａ It has the structure -L-, where L ａ It has a structure selected from the following group: 【Chemistry 2】 Furthermore, L has the structure shown below, where * represents L and L ａ It is the binding site, 【Transformation 3】 Group (B): 【Chemistry 4】 Group (c): 【Chemistry 5-1】 or 【Chemistry 5-2】 Here, the B ring and the C ring are each independently selected from the group consisting of C6 aryl compounds. Said J １ is -(Y) ｚ -, and said Y is selected from the group consisting of -C(O)-, -C(O)NH- Said J ２ is -(Y)z-, and each Y is selected from the group consisting of -C(O)- and -NHC(O)-, L ５ These are C1-C8 alkylenes, -NHC(O)-, or -C(O)NH-. L ６ is C １ -C ８ It is alkylene, -O-, or -C(O)NH-, Furthermore, each Y, L ５ , and L ６ As long as Y and L together form a chemically stable structure, ５ and L ６ R is optionally substituted with one or more Rs, and R is selected from the group consisting of H, -OH, and C1-C4 alkyl groups. m, n, s, and t are each independently selected from the group consisting of 0, 1, 2, 3, or 4. z is 1, The first drug molecule is a berberine derivative and is a drug molecule of the following formula II, formula III, or formula IV. 【Chemistry 6-1】 During the ceremony, Ro, Rp, Rq, Rr, Rs, and Rt are each independently selected from the group consisting of H, substituted or unsubstituted C1-C4 alkyl groups, and substituted or unsubstituted C1-C4 alkoxy groups, or Ro, Rp, Rq, Rr, Rs, and Rt are located on two adjacent atoms and together with the atoms linked to them form a 5-7 membered heterocycle, where substitution means that the H atom on the group is substituted with one or more substituents selected from the group consisting of halogens, C1-C4 alkyl groups, and phenyl groups. The second drug molecule is a JAK family inhibitor, and the JAK family inhibitors include tofacitinib, ruxolitinib, oclacitinib, baricitinib, peficitinib, abrocitinib, filgotinib, upadacitinib, and delgocitinib. The group consisting of ocitinib, itacitinib, fedratinib, decernotinib, SHR-0302, AZD-4205, ASN-002, BMS-986165, PF-06700841, PF-06651600, R-348, INCB-52793, ATI-501, ATI-502, NS-018, KL-130008, or a deuterated derivative of the above molecules, is selected. Co-pharmaceutical compounds.

2. The first drug molecule is a berberine analog, which is a drug molecule of the following formula III. 【Chemistry 6-2】 Ro, Rp, Rq, Rr, Rs, and Rt are each independently selected from the group consisting of H, substituted or unsubstituted C1-C4 alkyl groups, and substituted or unsubstituted C1-C4 alkoxy groups, or Ro, Rp, Rq, Rr, Rs, and Rt are located on two adjacent atoms and together with the atoms linked to them form a 5-7 membered heterocycle, where substitution means that the H atom on the group is substituted with one or more substituents selected from the group consisting of halogens, C1-C4 alkyl groups, and phenyl groups. The co-pharmaceutical compound according to claim 1.

3. The first drug group is one of the following groups 【Transformation 7】 Characterized by being selected from The co-pharmaceutical compound according to claim 1.

4. The first drug group is selected from the group consisting of the following: 【Chemistry 8-1】 Alternatively, the first drug group is a group formed by losing one hydrogen atom in the drug molecule, selected from the following group. 【Chemistry 8-2】 z is 1 The co-pharmaceutical compound according to claim 1.

5. The first drug group is given by the following formula 【Chemistry 9】 Characterized by having the structure shown. The co-pharmaceutical compound according to claim 1.

6. The second drug group is one of the following groups 【Chemistry 10】 【Chemistry 11】 Characterized by being selected from The co-pharmaceutical compound according to claim 1.

7. The second drug group is one of the following groups 【Chemistry 12】 Characterized by being selected from The co-pharmaceutical compound according to claim 1.

8. The aforementioned linkers belong to the following groups (A), (B), and (c): (A) Group is -L ａ It has the structure -L-, and the L ａ The structure is selected from the following group: 【Chemistry 13】 Furthermore, L has the structure shown below, where * represents L and L ａ This is the connection point, 【Chemistry 14】 Group (B): 【Chemistry 15】 Group (c): 【Chemistry 16】 Characterized by being selected from The co-pharmaceutical compound according to claim 1.

9. The following groups Table 1-1 Table 1-2 Table 1-3 Table 1-4 Characterized by being selected from The co-pharmaceutical compound according to claim 1.

10. A pharmaceutical composition, The pharmaceutical composition is characterized by comprising a therapeutically effective amount of the compound described in claim 1, or a stereoisomer or racemate thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

11. The pharmaceutical composition according to claim 10, characterized in that it is an enteric-coated preparation.

12. This treatment is characterized by its ability to treat diseases selected from the group consisting of gastrointestinal inflammatory disease, gastroenteritis caused by radiotherapy or chemotherapy, autoimmune diseases of the digestive system, peptic ulcers, irritable bowel syndrome, gastric cancer, esophageal cancer, and colon cancer. The pharmaceutical composition according to claim 10.

13. This treatment is characterized by its ability to treat diseases selected from the group consisting of ulcerative colitis, Crohn's disease, colitis associated with immune checkpoint inhibitor therapy, collagen colitis, lymphocytic colitis, pouchitis, acute / chronic gastritis, acute / chronic appendicitis, graft-versus-host disease, sprue, and autoimmune bowel diseases. The pharmaceutical composition according to claim 10.

14. The pharmaceutical composition according to claim 10, characterized in that it is used for the prevention and treatment of digestive system disorders.

15. The pharmaceutical composition according to claim 14, wherein the digestive functional disorder is a gastrointestinal inflammatory disease.

16. The aforementioned gastrointestinal inflammatory disease is characterized by being selected from the group consisting of ulcerative colitis, Crohn's disease, and colitis associated with immune checkpoint inhibitor therapy. The pharmaceutical composition according to claim 15.

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