CN107098876B

CN107098876B - Phenyl propionamide derivative, preparation method and medical application thereof

Info

Publication number: CN107098876B
Application number: CN201710095032.8A
Authority: CN
Inventors: 李心; 刘宪波; 贺峰; 陶维康; 孙飘扬
Original assignee: Jiangsu Hengrui Medicine Co Ltd; Shanghai Hengrui Pharmaceutical Co Ltd
Current assignee: Jiangsu Hengrui Medicine Co Ltd; Shanghai Hengrui Pharmaceutical Co Ltd
Priority date: 2016-02-23
Filing date: 2017-02-22
Publication date: 2021-04-06
Anticipated expiration: 2037-02-22
Also published as: CN107098876A

Abstract

The invention relates to phenyl propionamide derivatives, a preparation method and application thereof in medicines. In particular, the invention relates to phenylpropionamide derivatives represented by the general formula (I), a preparation method thereof, a pharmaceutical composition containing the derivatives, an application of the derivatives as kappa opioid receptor (KOR receptor) agonists, and an application of the derivatives in preparation of drugs for treating and/or preventing pain and pain-related diseases. Wherein each substituent group in the general formula (I) is defined as the specification.

Description

Phenyl propionamide derivative, preparation method and medical application thereof

Technical Field

The invention belongs to the field of medicines, and relates to a phenylpropionamide derivative, a preparation method thereof and application thereof in medicines. In particular, the invention relates to phenylpropionamide derivatives represented by the general formula (I), a preparation method thereof, a pharmaceutical composition containing the derivatives, and application of the derivatives as kappa opioid receptor (KOR receptor) agonists and application of the derivatives in preparation of drugs for treating and/or preventing pain and pain-related diseases.

Background

Opioid receptors are important G protein coupled receptors, are target points of combination of endogenous opioid peptides and opioid drugs, have a regulating effect on nervous system immunity and endocrine systems after being activated, and are the strongest and commonly used central analgesics at present. Endogenous opioid peptides are opioid active substances naturally produced in mammals, and currently known endogenous opioid peptides are roughly classified into enkephalins, endorphins, dynorphins and neokephalins (Pharmacol Rev 2007; 59: 88-123), and corresponding opioid receptors, namely mu, delta, kappa receptors and the like, exist in the central nervous system.

The Kappa Opioid Receptor (KOR) consists of 380 amino acids, and dynorphin is its endogenous ligand. It is expressed in sensory neurons, dorsal root ganglion cells and primary afferent neuron terminals, and participates in important physiological activities such as pain sensation, neuroendocrine, emotional behavior and cognition. It is now known that the human KOR is encoded by the OPRK1 gene and is located at chromosome 8q11-12 (Simonin F, Gaveriaux Ruff C, Kieffer BL, et al Proc Natl Acad Sci USA 1995,92(15): 7006-10). KOR activation and G protein G_i/G₀Coupling, increasing phosphodiesterase activity, inhibiting the activity of adenylate cyclase, reducing the intracellular cAMP level,thereby producing an inhibitory effect on neurons. KOR agonists have desensitizing effects on receptors with repeated action and reduced inhibition of adenylate cyclase activity (Raynor K, Kong H, Hines J, et al.J. Pharmacol Exp Ther,1994,270: 1381-6). KOR is also coupled to inward rectifying potassium channels and N-type calcium ion channels (Henry DJ, Grandy DK, Lester HA, Davidson N, Chavkin C (Mar 1995) Molecular Pharmacology 47(3): 551-7). KOR agonists are able to inhibit (calcium ion dependent) the release of pro-nociceptive and pro-inflammatory substance P from peripheral sensory nerve endings, which may be responsible for their antinociceptive and anti-inflammatory effects. In addition to dynorphin, various natural alkaloids and synthetic ligands can also bind to KOR. KORs provide natural addiction control mechanisms, and thus, drugs that are receptor agonists have potential as drug addiction treatments.

The effects of the KOR agonist asimadoline (asimadoline) in rodent diabetic neuropathy (Jolivalt et al. Diabetologia 2006,49(11): 2775-85; Epub Aug.19) and KOR agonist U-50488 in the rat Chronic Compressive Injury (CCI) model of neuropathic pain and the blockade of its effects by the opioid antagonist naloxone (naloxone) (Bileviciute-Ljungar et al. Eur.J. Pharm 2004.494:139-46) support the use of KOR agonists in the treatment of diabetic, viral and chemotherapy-induced neuropathic pain. The use of KOR agonists for the treatment or prevention of visceral pain including gynaecological disorders such as dysmenorrhoea spasticity and endometriosis has also been evaluated (Riviere, br.j. pharmacol 2004.141: 1331-4).

Kappa opioid agonists increase renal excretion of water and decrease natriuresis (i.e., produce selective water diuresis, also known as aquaresis), which many researchers believe is due to inhibition of pituitary secretion of antidiuretic hormones. Comparison of centrally acting and allegedly peripherally selective kappa opioids studies led to the conclusion that KOR within the blood brain barrier is responsible for mediating this effect. Researchers have proposed treating hyponatremia with nociceptin peptides or charged peptide conjugates that peripherally act on nociceptin receptors that are associated with, but not identical to, KORs (d.r.kapusta, Life sci.,60:15-21,1997).

Presently disclosed KOR agonist patent applications include WO20071398, WO2008060552, WO09932510, WO2013184794, WO2014089019, WO2014184356, and WO 2015065867.

The kappa opioid receptor (KOR receptor) agonist has a good application prospect in the pharmaceutical industry as a medicine, and in order to achieve the purpose of better treatment effect and better meet the market demand, the inventor hopes to develop a new generation of KOR agonist with high efficiency and low toxicity. The present invention is to provide a novel kappa opioid receptor (KOR receptor) agonist compound which surprisingly exhibits excellent effects and effects.

Disclosure of Invention

The invention aims to provide a compound shown in a general formula (I):

or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof,

wherein:

g is C ═ O or O ═ S ═ O;

R¹selected from the group consisting of hydrogen, alkyl, alkoxy, haloalkyl, halogen, amino, nitro, hydroxy, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR³、-C(O)R³、-C(O)OR³、-S(O)_mR³and-NR⁴R⁵Wherein said alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally selected from the group consisting of alkyl, haloalkyl, halogen, amino, nitro, cyano, hydroxy, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and NR⁶R⁷Is substituted with one or more substituents of (1);

R²selected from halogen, alkoxy, haloalkyl, -OR³Cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein said alkyl, cycloalkyl, heteroarylThe cyclic, aryl and heteroaryl groups are optionally substituted with one or more substituents selected from the group consisting of halogen, amino, nitro, cyano, hydroxy, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

R³selected from the group consisting of hydrogen atoms, alkyl groups, amino groups, alkoxy groups, hydroxyalkyl groups, cycloalkyl groups, heterocyclyl groups, aryl groups, and heteroaryl groups, wherein said alkyl groups, cycloalkyl groups, heterocyclyl groups, aryl groups, and heteroaryl groups are optionally substituted with one or more substituents selected from the group consisting of alkyl groups, halogen groups, hydroxy groups, amino groups, nitro groups, cyano groups, alkoxy groups, hydroxyalkyl groups, cycloalkyl groups, heterocyclyl groups, aryl groups, and heteroaryl groups;

R⁴and R⁵Each independently selected from the group consisting of hydrogen atoms, alkyl groups, alkoxy groups, hydroxyalkyl groups, hydroxyl groups, amino groups, carboxylate groups, cycloalkyl groups, heterocyclyl groups, aryl groups, and heteroaryl groups, wherein said alkyl groups, cycloalkyl groups, heterocyclyl groups, aryl groups, and heteroaryl groups are optionally substituted with one or more substituents selected from the group consisting of alkyl groups, halogen groups, hydroxyl groups, amino groups, carboxylate groups, nitro groups, cyano groups, alkoxy groups, hydroxyalkyl groups, cycloalkyl groups, heterocyclyl groups, aryl groups, and heteroaryl groups;

R⁶and R⁷Each independently is a hydrogen atom or W;

w is an amino protecting group;

m is 0, 1 or 2; and is

n is 1,2, 3 or 4.

In a preferred embodiment of the present invention, the compound represented by the general formula (I) is a compound represented by the general formula (II):

wherein:

R¹、R²and n is as defined in formula (I).

In a preferred embodiment of the present invention, the compound represented by the general formula (I) is a compound represented by the general formula (III):

wherein:

G、R²and n is as defined in formula (I).

In a preferred embodiment of the present invention, the compounds of the general formulae (I) to (III) wherein R is²Selected from halogen, OR³Aryl and heterocyclyl, wherein said aryl and heterocyclyl are optionally substituted by one or more alkyl or halogen, R³Is an alkyl group.

In a preferred embodiment of the invention, the compounds of formula (I) and formula (III) wherein R is¹Is alkyl, wherein said alkyl is optionally substituted with NR⁶R⁷Substituted by a substituent; r⁶And R⁷Each independently is a hydrogen atom or W; w is an amino protecting group.

Typical compounds of formula (I) include, but are not limited to:

or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

The present invention further provides a compound of formula (IV) as an intermediate for the preparation of formula (I):

wherein:

w is an amino protecting group;

G、R¹、R²and n is as defined in formula (I).

Compounds of formula (IV) include, but are not limited to:

the present invention further provides a process for the preparation of a compound according to general formula (I) or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, comprising:

reacting the compound of the general formula (I-4) with the compound of the general formula (I-5) to obtain a compound of a general formula (IV); further removing the protecting group of the obtained compound of the general formula (IV) to obtain a compound of a general formula (I);

wherein:

w is an amino protecting group;

G、n、R¹and R²As defined in formula (I).

Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), (II), (III) above, or a tautomer, mesomer, racemate, enantiomer, or diastereomer thereof, or mixture thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients. The invention also relates to a method for preparing the pharmaceutical composition, which comprises mixing the compound shown in the general formula (I), (II) and (III) or the tautomer, the mesomer, the racemate, the enantiomer, the diastereomer or the mixture form or the pharmaceutically acceptable salt form thereof with a pharmaceutically acceptable carrier, diluent or excipient.

In one embodiment, the pharmaceutical composition of the present invention further comprises one or more of the following compounds: opioids, cannabinoids, antidepressants, anticonvulsants, tranquilizers, corticosteroids, ion channel blockers or non-steroidal anti-inflammatory drugs (NSAIDs).

The invention further relates to the use of a compound of general formula (I), (II), (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for the preparation of a medicament for agonizing or antagonizing the kappa opioid receptor (KOR receptor).

The invention further relates to the use of a compound of general formula (I), (II), (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for the manufacture of a medicament for the prevention and/or treatment of a kappa opioid receptor (KOR receptor) agonist-mediated related disorder, preferably selected from pain, inflammation, pruritus, edema, hyponatremia, hypokalemia, ileus, cough and glaucoma, more preferably pain.

The invention further relates to application of the compound shown in the general formula (I), (II), (III) or a tautomer, a mesomer, a racemate, an enantiomer, a diastereomer or a mixture form thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the compound in preparation of medicines for preventing and/or treating pain and pain related diseases of mammals (such as human beings), wherein the pain can be postoperative pain, cancer-caused pain, neuropathic pain, traumatic pain, inflammation-caused pain and the like.

The present invention further relates to a method of agonizing or antagonizing the kappa opioid receptor (KOR receptor) comprising administering to a patient in need thereof a therapeutically effective dose of a compound of the present invention of formula (I), (II), (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof.

The present invention further relates to a method for the prevention and/or treatment of kappa opioid receptor (KOR receptor) agonist receptor-mediated disorders comprising administering to a patient in need thereof a therapeutically effective dose of a compound of the present invention of the general formula (I), (II), (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof. The method has remarkable therapeutic effect and less side effects. Wherein said kappa opioid receptor (KOR receptor) agonist receptor-mediated related disorder is selected from the group consisting of pain, inflammation, itch, edema, hyponatremia, hypokalemia, ileus, cough and glaucoma, preferably pain.

The invention further relates to a compound of the general formula (I), (II), (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, for use as a medicament.

The invention further relates to compounds of the general formula (I), (II), (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, for use in agonizing or antagonizing the kappa opioid receptor (KOR receptor).

The invention further relates to compounds of the general formula (I), (II), (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, for use in the prevention and/or treatment of a kappa opioid receptor (KOR receptor) receptor agonist-mediated related disease.

The invention further relates to compounds of general formula (I), (II), (III) or tautomers, mesomers, racemates, enantiomers, diastereomers or mixtures thereof, or pharmaceutically acceptable salts thereof, which are useful for the prevention and/or treatment of pain and pain-related disorders in mammals (e.g., humans).

The kappa opioid receptor agonist-mediated associated disease, disorder or condition may be any kappa opioid receptor (KOR receptor) agonist-mediated associated condition, including but not limited to acute or chronic pain, inflammation, itch, hyponatremia, edema, ileus, cough, and glaucoma. For example, kappa opioid receptor (KOR receptor) associated pain may be neuropathic pain, somatic pain, visceral pain, or skin pain. Some diseases, disorders or conditions are associated with more than one form of pain. For example, the post-operative pain may be any or all of neuropathic, somatic, visceral or cutaneous pain factors, depending on the type and extent of surgery employed.

The kappa opioid receptor (KOR receptor) associated inflammation to which the present invention relates may be any inflammatory disease or condition, including, but not limited to, sinusitis, rheumatoid arthritis tenosynovitis, bursitis, tendonitis, external humeral epicondylitis, bursitis, osteomyelitis, osteoarthritis, Inflammatory Bowel Disease (IBD), Irritable Bowel Syndrome (IBS), ocular inflammation, otic inflammation, or autoimmune inflammation.

The kappa opioid receptor (KOR receptor) -associated itch referred to herein may be any itch disease and condition, for example ocular itch such as conjunctivitis-associated ocular itch, end-stage kidney disease-associated itch in which many patients receive kidney dialysis, and other forms of cholestasis including primary biliary cirrhosis, intrahepatic cholestasis of pregnancy, chronic cholesterol liver disease, uremia, malignant cholestasis, jaundice, and skin conditions such as eczema (dermatitis) including atopic dermatitis or contact dermatitis, dermatophytosis, erythrocytosis, lichen planus, chronic lichen simplex, pediculosis, thyrotoxicosis, tinea pedis, urticaria, scabies, vaginitis, hemorrhoid-associated anal itch, and insect bite itch and drug-induced itch, for example mu opioid-induced itch.

The kappa opioid receptor (KOR receptor) -associated edema to which the present invention relates may be any edematous disease or condition, such as edema caused by congestive heart disease or edema caused by the syndrome of inappropriate secretion of antidiuretic hormone (ADH).

Kappa opioid receptor (KOR receptor) -associated ileus contemplated by the present invention may be any ileus disease or condition, including but not limited to post-operative ileus or opioid-induced bowel dysfunction.

The kappa opioid receptor (KOR receptor) -associated neuropathic pain to which the present invention relates may be any neuropathic pain, for example, trigeminal neuralgia, diabetic pain, viral-induced pain such as herpes zoster-associated pain, chemotherapy-induced pain, metastatic cancer pain of the affected nerve, traumatic and surgical-associated neuropathic pain, and various headache variants believed to have neuropathological factors such as migraine.

The kappa opioid receptor (KOR receptor) -associated pain cough related to the present invention includes ocular pain such as ocular pain after photorefractive keratectomy (PRK), ocular laceration, ocular fundus fracture, chemical burn, corneal epithelial abrasion or irritation, or ocular pain associated with conjunctivitis, corneal ulcer, scleritis, episcleritis, scleral keratitis, ocular herpes zoster, interstitial keratitis, acute iritis, keratoconjunctivitis sicca, orbital cellulitis, orbital pseudotumor, pemphigus, trachoma or uveitis.

The pain associated with the kappa opioid receptor (KOR receptor) to which the present invention relates also includes sore throat, particularly associated with inflammatory conditions such as allergic rhinitis, acute bronchitis, the common cold, contact ulcers, herpes simplex virus injury, infectious mononucleosis, influenza, laryngeal cancer, acute laryngitis, acute necrotizing ulcerative gingivitis, tonsillitis abscess, pharyngeal burning, pharyngitis, reflux pharyngolaryngitis, acute sinusitis, and tonsillitis.

The pain associated with the kappa opioid receptor (KOR receptor) to which the present invention relates may be arthritic pain, kidney stones, urinary and biliary calculi pain, uterine cramps, dysmenorrhea, endometriosis, mastitis, dyspepsia, post-surgical pain (e.g., post-appendectomy, open colorectal surgery, hernia repair, prostatectomy, colectomy, gastrectomy, splenectomy, colectomy, colostomy, pelvic laparoscopic examination, tubal ligation, hysterectomy, vasectomy, or cholecystectomy-induced post-surgical pain), post-medical treatment pain (e.g., pain following colonoscopy, cystoscopy, hysteroscopy, or cervical or endometrial biopsy), otitis pain, breakthrough cancer pain, and pain associated with GI disorders such as IBD or IBS or other inflammatory conditions, in particular pain associated with visceral inflammation (e.g. gastroesophageal reflux disease, pancreatitis, acute pyelonephritis, ulcerative colitis, cholecystitis, cirrhosis, liver cysts, hepatitis, duodenal or gastric ulcers, esophagitis, gastritis, gastroenteritis, colitis, diverticulitis, intestinal obstruction, ovarian cysts, pelvic inflammatory disease, perforation of ulcers, peritonitis, prostatitis, interstitial cystitis), or pain from contact with toxic agents (e.g. insect toxins, or drugs such as salicylates or NSAIDs).

The kappa opioid receptor (KOR receptor) associated hyponatremia to which the present invention relates may be any disease or condition in which hyponatremia (a low sodium condition) exists, for example, in humans, abnormalities may occur alone when the concentration of sodium in the plasma is below 135mmol/l, or more commonly as a complication of other medical conditions or as a result of the use of drugs that may cause sodium deficiency, wherein said hyponatremia-related diseases include, but are not limited to: neoplastic factors that cause excessive ADH secretion, including cancers of the lung, duodenum, pancreas, ovary, bladder, and ureter, thymoma, mesothelioma, bronchial adenoma, carcinoid tumors, ganglioneuroma, and ewing's sarcoma, infection; such as pneumonia (bacterial or viral), abscesses (lung or brain), vacuolization (aspergillosis), tuberculosis (lung or brain), meningitis (bacterial or viral), encephalitis, and AIDS; vascular factors such as: cerebral infarction or hemorrhage and cavernous sinus embolism; neurological factors such as: Guillan-Barre syndrome, multiple sclerosis, tremor delirium, amyotrophic lateral sclerosis, hydrocephalus, psychosis, peripheral neuropathy, head trauma (occlusive and penetrating), CNS tumors or infections and CNS lesions affecting hypothalamic osmoreceptors; congenital malformation involving the whole life: corpus callosum hypoplasia, cleft lip and palate and other midline defects; metabolic factors such as: acute intermittent porphyria, asthma, pneumothorax, and positive pressure breathing; drugs such as: thiazine diuretics, paracetamol, barbiturates, cholinomimetics, estrogens, oral hypoglycemic agents, vasopressin or desmopressin, high-dose oxytocin chlorpropamide, vincristine, carbamazepine, nicotine, phenothiazine, cyclophosphamide, tricyclic antidepressants, monoamine oxidase inhibitors, and 5-hydroxytryptamine reuptake inhibitors; such as the administration of excess hypotonic fluids during or after hospitalization, surgery or physical activity (i.e., exercise-related hyponatremia), and the use of low-sodium nutritional supplements in elderly individuals, other conditions associated with hyponatremia include renal failure, nephrotic syndrome (model nephropathy and minimal disease), malignancy, malnutrition, rhabdomyolysis, surgical management, selective cardiac catheterization, blood loss, and hypercalcemia, hypokalemia, and hyperglycemia whose consequences are diabetes that can lead to osmotic diuresis.

Another aspect of the present invention relates to a method for the prevention and/or treatment of a kappa opioid receptor (KOR receptor) agonist-mediated related disease, disorder or condition-related disease, comprising administering to a patient in need thereof a therapeutically effective dose of a compound of the present invention, in particular of formula (I) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof; the method exhibits outstanding therapeutic effects and fewer side effects, wherein the kappa opioid receptor (KOR receptor) agonist-mediated related disorders include, but are not limited to, acute or chronic pain, inflammation, pruritus, hyponatremia, edema, ileus, cough and glaucoma.

Another aspect of the present invention relates to a method for preventing and/or treating pain and pain-related diseases in a mammal, which comprises administering to a mammal in need thereof a therapeutically effective amount of a compound of the present invention represented by the general formula (I), (II), (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof; the method exhibits outstanding therapeutic efficacy and fewer side effects, wherein the pain may be post-operative pain, cancer-induced pain, neuropathic pain, traumatic pain, somatic pain, visceral pain, skin pain and inflammation-induced pain, e.g. post-operative pain may be any or all of neuropathic pain, somatic pain, visceral pain or skin pain factors, depending on the type and extent of the surgery employed; the cancer may be selected from breast cancer, endometrial cancer, cervical cancer, skin cancer, prostate cancer, ovarian cancer, fallopian tube tumor, ovarian tumor, hemophilia, leukemia, etc.

The pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Oral compositions may be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more ingredients selected from the group consisting of: sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide a pleasant to the eye and palatable pharmaceutical preparation. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be inert excipients, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as microcrystalline cellulose, croscarmellose sodium, corn starch or alginic acid; binding agents, for example starch, gelatin, polyvinylpyrrolidone or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc. These tablets may be uncoated or they may be coated by known techniques which mask the taste of the drug or delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, water soluble taste masking substances such as hydroxypropylmethyl cellulose or hydroxypropyl cellulose, or time extending substances such as ethyl cellulose, cellulose acetate butyrate may be used.

Oral formulations may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with a water soluble carrier, for example polyethylene glycol, or an oil vehicle, for example peanut oil, liquid paraffin or olive oil.

Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone and acacia; dispersing or wetting agents may be a naturally-occurring phosphatide, for example lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyethylene oxide sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene oxide sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl paraben, one or more colouring agents, one or more flavouring agents and one or more sweetening agents, such as sucrose, saccharin or aspartame.

Oil suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oil suspension may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable preparation. These compositions can be preserved by the addition of antioxidants such as butylated hydroxyanisole or alpha-tocopherol.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent or one or more preservatives. Suitable dispersing or wetting agents and suspending agents are illustrative of the examples given above. Other excipients, for example sweetening, flavoring and coloring agents, may also be present. These compositions are preserved by the addition of an antioxidant such as ascorbic acid.

The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin or mixtures thereof. Suitable emulsifying agents may be naturally-occurring phosphatides, for example soy bean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyethylene oxide sorbitol monooleate. The emulsions may also contain sweetening agents, flavouring agents, preservatives and antioxidants. Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, a colorant and an antioxidant.

The pharmaceutical compositions of the present invention may be in the form of a sterile injectable aqueous solution. Among the acceptable vehicles or solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oil phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. The oil solution is then treated to form a microemulsion by adding to a mixture of water and glycerol. The injection solution or microemulsion may be injected into the bloodstream of a patient by local bulk injection. Alternatively, it may be desirable to administer the solutions and microemulsions in a manner that maintains a constant circulating concentration of the compounds of the present invention. To maintain such a constant concentration, a continuous intravenous delivery device may be used. An example of such a device is an intravenous pump model Deltec CADD-PLUS. TM.5400.

The pharmaceutical compositions of the present invention may be in the form of sterile injectable aqueous or oleaginous suspensions for intramuscular and subcutaneous administration. The suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension prepared in a parenterally-acceptable non-toxic diluent or solvent, for example as a solution in 1, 3-butanediol. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any blend fixed oil may be used, including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The compounds of the present invention may be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid in the rectum and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, glycerogelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights and mixtures of fatty acid esters of polyethylene glycols.

As is well known to those skilled in the art, the dosage of a drug administered depends on a variety of factors, including, but not limited to: the activity of the particular compound employed, the age of the patient, the weight of the patient, the health condition of the patient, the patient's integument, the patient's diet, the time of administration, the mode of administration, the rate of excretion, the combination of drugs, and the like; in addition, the optimal treatment regimen, such as the mode of treatment, the daily amount of compound (I) of the formula or the type of pharmaceutically acceptable salt, can be verified according to conventional treatment protocols.

Detailed description of the invention

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

The term "alkyl" refers to a saturated aliphatic hydrocarbon group which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-dimethylpentyl, 2, 4-dimethylpentyl, 2-dimethylpentyl, 3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2, 3-dimethylhexyl, 2, 4-dimethylhexyl, 2, 5-dimethylhexyl, 2-dimethylhexyl, 3-dimethylhexyl, 4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2-dimethylpentyl, 2-dimethylhexyl, 3-dimethylpentyl, 2-ethylhexyl, 3-dimethylhexyl, 2, 2-diethylpentyl, n-decyl, 3-diethylhexyl, 2-diethylhexyl, and various branched isomers thereof. More preferred are lower alkyl groups having 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl and the like. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halo, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring containing from 3 to 20 carbon atoms, preferably from 3 to 12 carbon atoms, more preferably from 3 to 6 carbon atoms, and most preferably from 5 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl groups.

The term "spirocycloalkyl" refers to a 5 to 20 membered polycyclic group sharing one carbon atom (referred to as a spiro atom) between monocyclic rings, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. Spirocycloalkyl groups are classified into a single spirocycloalkyl group, a double spirocycloalkyl group or a multi spirocycloalkyl group, preferably a single spirocycloalkyl group and a double spirocycloalkyl group, according to the number of spiro atoms shared between rings. More preferably 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered. Non-limiting examples of spirocycloalkyl groups include:

the term "fused cyclic alkyl" refers to a 5 to 20 membered all carbon polycyclic group in which each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, wherein one or more of the rings may contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyls according to the number of constituent rings, preferably bicyclic or tricyclic, more preferably 5-or 6-membered bicycloalkyl. Non-limiting examples of fused ring alkyl groups include:

the term "bridged cycloalkyl" refers to a 5 to 20 membered all carbon polycyclic group in which any two rings share two carbon atoms not directly attached, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups according to the number of constituent rings, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl groups include:

the cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, where the ring to which the parent structure is attached is cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, and the like; preferably phenyl and cyclopentyl, tetrahydronaphthyl. Cycloalkyl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "heterocyclyl" refers to a saturated or partially unsaturated mono-or polycyclic cyclic hydrocarbon substituent containing from 3 to 20 ring atoms wherein one or more of the ring atoms is selected from nitrogen, oxygen, or S (O)_m(wherein m is an integer from 0 to 2) but excludes the ring moiety of-O-O-, -O-S-, or-S-S-, the remaining ring atoms being carbon. Preferably 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; most preferably 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; most preferably 5 to 6 ring atoms, of which 1-2 or 1-3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like, with tetrahydropyranyl, piperidinyl, pyrrolidinyl being preferred. Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups.

The term "spiroheterocyclyl" refers to a 5 to 20 membered monocyclic ring which shares one ring with anotherPolycyclic heterocyclic radicals of atoms, known as spiro atoms, in which one or more ring members are selected from nitrogen, oxygen or S (O)_m(wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. The spiro heterocyclic group is classified into a mono-spiro heterocyclic group, a di-spiro heterocyclic group or a multi-spiro heterocyclic group, preferably a mono-spiro heterocyclic group and a di-spiro heterocyclic group, according to the number of spiro atoms shared between rings. More preferred are 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered mono spiroheterocyclic groups. Non-limiting examples of spiro heterocyclic groups include:

the term "fused heterocyclyl" refers to a 5 to 20 membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system, one or more rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system in which one or more ring atoms is selected from nitrogen, oxygen or S (O)_m(wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups according to the number of constituent rings, preferably bicyclic or tricyclic, more preferably 5-or 6-membered bicyclic fused heterocyclic groups. Non-limiting examples of fused heterocyclic groups include:

the term "bridged heterocyclyl" refers to a 5 to 14 membered polycyclic heterocyclic group in which any two rings share two atoms not directly attached which may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system in which one or more of the ring atoms is selected from nitrogen, oxygen or S (O)_m(wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14Preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups according to the number of constituent rings, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

the heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring to which the parent structure is attached is heterocyclyl, non-limiting examples of which include:

and the like.

The heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "aryl" refers to a 6 to 14 membered all carbon monocyclic or fused polycyclic (i.e., rings which share adjacent pairs of carbon atoms) group having a conjugated pi-electron system, preferably 6 to 10 membered, more preferably 5 to 6 membered, such as phenyl and naphthyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring, non-limiting examples of which include:

the aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy or carboxylate.

The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5 to 10 membered, containing 1 to 3 heteroatoms; more preferably 5 or 6 membered, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl and the like, preferably imidazolyl, pyrazolyl or pyrimidinyl, thiazolyl; more preferably pyrazolyl. The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring joined together with the parent structure is a heteroaryl ring, non-limiting examples of which include:

heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate groups.

The term "alkoxy" refers to-O- (alkyl) and-O- (unsubstituted cycloalkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy or carboxylate groups.

The term "amino protecting group" refers to a group suitable for protecting (preventing) an amino group from chemical reaction, but which is easily removed after the chemical reaction to be performed elsewhere in the molecule is completed. These groups typically represent unsubstituted or substituted acyl groups, unsubstituted or substituted allyl groups, aryl groups, aralkyloxymethyl groups, aralkyl groups or together with the nitrogen atom form heterocyclic groups and salts. Non-limiting examples include t-butyloxycarbonyl (Boc), benzyloxycarbonyl, isobutyloxycarbonyl, fluorenylmethyloxycarbonyl (Fmoc), benzoyl, substituted benzoyl, butyryl, acetyl, trifluoroacetyl, phthalimido (Pht), succinimidyl, maleimido, benzyl, allyloxycarbonyl, and p-methoxybenzyl, and the like. These groups may be optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, hydroxy, nitro, acylamino, acyl, and the like substituted benzyl, o-methylbenzyl, trityl, and benzhydryl. The amino protecting groups are preferably t-butyloxycarbonyl and fluorenylmethyloxycarbonyl (Fmoc).

The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.

The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

The term "hydroxy" refers to an-OH group.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "amino" refers to the group-NH₂。

The term "cyano" refers to — CN.

The term "nitro" means-NO₂。

The term "carboxy" refers to-C (O) OH.

The term "carboxylate" refers to-C (O) O (alkyl) or-C (O) O (cycloalkyl), wherein alkyl, cycloalkyl are as defined above.

The term "acyl halide" refers to a compound containing a group that is-C (O) -halogen.

Different terms such as "X is selected from A, B or C", "X is selected from A, B and C", "X is A, B or C", "X is A, B and C" and the like all express the same meaning, that is, X can be any one or more of A, B, C.

"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not. For example, "a heterocyclic group optionally substituted with an alkyl" means that an alkyl may, but need not, be present, and the description includes the case where the heterocyclic group is substituted with an alkyl and the heterocyclic group is not substituted with an alkyl.

"substituted" means that one or more, preferably up to 5, more preferably 1 to 3, hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable in combination with carbon atoms having unsaturated (e.g., olefinic) bonds.

"pharmaceutical composition" means a mixture containing one or more compounds described herein or a physiologically/pharmaceutically acceptable salt or prodrug thereof in admixture with other chemical components, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of the active ingredient and exert biological activity.

"pharmaceutically acceptable salts" refers to salts of the compounds of the present invention which are safe and effective for use in the body of a mammal and which possess the requisite biological activity.

Synthesis of the Compounds of the invention

In order to achieve the purpose of the invention, the invention adopts the following technical scheme:

the invention relates to a method for preparing a compound shown in a general formula (I) or a tautomer, a mesomer, a racemate, an enantiomer, a diastereomer or a mixture form thereof, or a pharmaceutically acceptable salt form thereof, which comprises the following steps:

reacting the compound of the general formula (I-1) or the salt thereof with the compound of the general formula (I-2) under alkaline conditions to obtain a compound of the general formula (I-3); removing the protecting group on the amino group of the obtained compound of the general formula (I-3) under alkaline conditions to obtain a compound of the general formula (I-4) or a salt thereof; carrying out condensation reaction on the obtained compound of the general formula (I-4) or salt thereof and the compound of the general formula (I-5) under alkaline conditions to obtain a compound of a general formula (IV); further removing the protecting group of the obtained general formula (IV) under an acidic condition to obtain a compound of the general formula (I) or a salt thereof;

the reagents that provide basic conditions include organic bases including, but not limited to, piperidine, triethylamine, N-diisopropylethylamine, N-butyllithium, lithium diisopropylamide, potassium acetate, sodium tert-butoxide, or potassium tert-butoxide, and inorganic bases including, but not limited to, sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, and lithium hydroxide.

Reagents that provide acidic conditions include, but are not limited to, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, and methanesulfonic acid.

Condensing agents include, but are not limited to, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, N, N '-dicyclohexylcarbodiimide, N, N' -diisopropylcarbodiimide, O-benzotriazol-N, N, N ', N' -tetramethyluronium tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazol, O-benzotriazol-N, N, N ', N' -tetramethyluronium hexafluorophosphate, 2- (7-oxybenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate, 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate, Benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate or benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate, preferably 2- (7-benzotriazol-oxide) -N, N' -tetramethyluronium hexafluorophosphate.

The above reaction is preferably carried out in a solvent including, but not limited to: acetic acid, methanol, ethanol, toluene, tetrahydrofuran, dichloromethane, dimethyl sulfoxide, 1, 4-dioxane, water or N, N-dimethylformamide.

Wherein:

w is an amino protecting group;

G、n、R¹and R²As defined in formula (I).

Detailed Description

The present invention is further described below with reference to examples, which are not intended to limit the scope of the present invention.

Examples

The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) or/and Mass Spectrometry (MS). NMR shift (. delta.) of 10^-6The units in (ppm) are given. NMR was measured using a Bruker AVANCE-400 NMR spectrometer using deuterated dimethyl sulfoxide (DMSO-d)₆) Deuterated chloroform (CDCl)₃) Deuterated methanol (CD)₃OD), internal standard Tetramethylsilane (TMS).

MS was determined using a FINNIGAN LCQAD (ESI) mass spectrometer (manufacturer: Thermo, model: Finnigan LCQ advantage MAX).

HPLC was carried out using an Agilent 1200DAD high pressure liquid chromatograph (Sunfire C18150X 4.6mm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18150X 4.6mm column).

Average inhibition rate of kinase and IC₅₀The values were determined with a NovoStar microplate reader (BMG, Germany).

The thin layer chromatography silica gel plate adopts HSGF254 of tobacco yellow sea or GF254 of Qingdao, the specification of the silica gel plate used by Thin Layer Chromatography (TLC) is 0.15 mm-0.2 mm, and the specification of the thin layer chromatography separation and purification product is 0.4 mm-0.5 mm.

The column chromatography generally uses 200-300 mesh silica gel of the Tibet Huanghai silica gel as a carrier.

Known starting materials of the present invention may be synthesized by or according to methods known in the art, or may be purchased from companies such as ABCR GmbH & Co.KG, Acros Organics, Aldrich Chemical Company, Shao Yuan Chemical technology (Accela ChemBio Inc), Darri Chemicals, and the like.

In the examples, the reaction can be carried out in an argon atmosphere or a nitrogen atmosphere, unless otherwise specified.

An argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to a balloon of argon or nitrogen with a volume of about 1L.

The hydrogen atmosphere refers to a reaction flask connected with a hydrogen balloon with a volume of about 1L.

The pressure hydrogenation reaction used a hydrogenation apparatus of Parr 3916EKX type and a hydrogen generator of Qinglan QL-500 type or a hydrogenation apparatus of HC2-SS type.

The hydrogenation reaction was usually evacuated and charged with hydrogen and repeated 3 times.

The microwave reaction was carried out using a CEM Discover-S908860 type microwave reactor.

In the examples, the solution means an aqueous solution unless otherwise specified.

In the examples, the reaction temperature is, unless otherwise specified, from 20 ℃ to 30 ℃ at room temperature.

The progress of the reaction in the examples was monitored by Thin Layer Chromatography (TLC) using a developing solvent system of: a: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: petroleum ether and ethyl acetate system, D: the volume ratio of acetone and solvent is adjusted according to the polarity of the compound.

The eluent system for column chromatography and the developing agent system for thin-layer chromatography used for purifying compounds comprise: a: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of basic or acidic reagents such as triethylamine, acetic acid and the like can be added for adjustment.

Example 1

4- ((R) -6-amino-2- ((R) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanoylamino) hexanoyl) -N- ((tetrahydro-2H-pyran-4-yl) methyl) piperazine-1-carboxamide 1

First step of

((tetrahydro-2H-pyran-4-yl) methyl) carbamic acid phenyl ester 1c

(tetrahydro-2H-pyran-4-yl) methylamine 1a (0.5g, 4.35mmol) and pyridine (0.41g, 5.22mmol) were dissolved in 15mL of tetrahydrofuran, and phenyl chloroformate 1b (750mg, 4.78mmol) was added dropwise at 0 ℃ to raise the temperature of the reaction mixture to room temperature, followed by stirring for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate, washed with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the crude title product 1c (1.1g), which was subjected to the next reaction without purification.

Second step of

4- (((tetrahydro-2H-pyran-4-yl) methyl) carbamoyl) piperazine-1-carboxylic acid benzyl ester 1e

Crude 1c (1.1g, 4.35mmol), piperazine-1-carboxylic acid benzyl ester 1d (1g, 4.54mmol) was dissolved in 20mL of methanol and the reaction was stirred at 50 ℃ for 12 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 1e (0.88g, yield: 56%).

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

The third step

N- ((tetrahydro-2H-pyran-4-yl) methyl) piperazine-1-carboxamide 1f

1e (0.15g, 0.415mmol) was dissolved in 10mL of methanol, palladium on carbon (0.1g, 10%) was added, and the reaction mixture was replaced with hydrogen three times, followed by stirring for 1 hour. The reaction was filtered and the filtrate was concentrated under reduced pressure to give the crude title product 1f (0.1g) which was directly subjected to the next reaction without purification.

MS m/z(ESI):228.2[M+1]

The fourth step

(R) - (9H-fluoren-9-yl) methyl tert-butyl (6-oxo-6- (4- (((tetrahydro-2H-pyran-4-yl) methyl) carbamoyl) piperazin-1-yl) hexane-1, 5-diyl) dicarbamate 1H

Crude 1f (0.1g, 0.415mmol), (R) -2- ((((9H-fluoren-9-yl) methoxy) carbonyl) amino) -6- ((tert-butoxycarbonyl) amino) hexanoic acid 1g (0.2g, 0.415mmol), prepared by a known method "Tetrahedron, 2002,58(27), 5427-5439"), was dissolved in 5mL of N, N-dimethylformamide, and 2- (7-azobenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (0.236g, 0.623mmol), triethylamine (0.11mL, 0.83mmol) and stirred for 12 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 1h (135mg, yield: 49%).

MS m/z(ESI):678.4[M+1]

The fifth step

(R) - (5-amino-6-oxo-6- (4- (((tetrahydro-2H-pyran-4-yl) methyl) carbamoyl) piperazin-1-yl) hexyl) carbamic acid tert-butyl ester 1j

1h (135mg, 0.202mmol) was dissolved in 5mL of dichloromethane, 0.5mL of piperidine was added, and the reaction was stirred for 4 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give 1j (92mg, yield: 100%).

MS m/z(ESI):456.3[M+1]

The sixth step

N- [ (5R) -5- [ (2R) -2- [ (2R) -2- [ (2R) -2- { [ (tert-butoxy) carbonyl ] amino } -3-phenylpropionamino ] -4-methylpentanylamino ] -6- {4- [ (tetrahydro-2H-pyran-4-ylmethyl) carbamoyl ] piperazin-1-yl } -6-oxohexyl ] carbamic acid tert-butyl ester 1m

1j (92mg, 0.202mmol), (6R,9R,12R) -6, 9-dibenzyl-12-isobutyl-2, 2-dimethyl-4, 7, 10-trioxo-3-oxa-5, 8, 11-triazatridecyl-13-carboxylic acid 1k (106mg, 0.202mmol, prepared by the method disclosed in patent application "US 20110212882"), 2- (7-azobenzenetriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (115mg, 0.303mmol) and triethylamine (0.056mL, 0.404mmol) were dissolved in 5mL of N, N-dimethylformamide and stirred for 12 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 1m (50mg, yield: 26%).

MS m/z(ESI):964.4[M+1]

Seventh step

1m (50mg, 0.052mmol) was dissolved in 5mL of dichloromethane, 0.5mL of trifluoroacetic acid was added, and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by high performance liquid chromatography to give the title product 1(4mg, yield: 10%).

MS m/z(ESI):763.4[M+1]

¹H NMR(400MHz,DMSO-d₆):δ8.41(d,1H),8.13(d,1H),7.38-7.28(m,10H),7.20(d,1H),6.05(s,1H),5.16(s,4H),4.69(d,1H),4.41-4.35(m,2H),4.11(d,1H),3.71(d,1H),3.63(d,2H),3.51(d,2H),3.27-3.23(m,3H),3.11-2.95(m,6H),2.20(d,1H),2.16-1.91(m,2H),1.85-1.56(m,7H),1.51-1.23(m,10H),1.01(d,3H),0.97(d,3H).

Example 2

4- ((R) -6-amino-2- ((R) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanoylamino) hexanoyl) -N- (4-fluorobenzyl) piperazine-1-carboxamide 2

First step of

(R) - (9H-fluoren-9-yl) methyl tert-butyl (6- (4- ((4-fluorobenzyl) carbamoyl) piperazin-1-yl) -6-oxohexane-1, 5-diyl) dicarbamate 5b

1g (468mg, 1mmol) was dissolved in 15mL of N, N-dimethylformamide, and N- (4-fluorobenzyl) piperazine-1-carboxamide hydrochloride 2a (273mg, 1mmol, prepared by the method disclosed in patent application "WO 2014039714"), 2- (7-azobenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (456mg, 1.2mmol), and N, N-diisopropylethylamine (0.3mL, 1.5mmol) were added and reacted for 12 hours with stirring. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 2b (600mg, yield: 87.2%).

Second step of

(R) - (5-amino-6- (4- ((4-fluorobenzyl) carbamoyl) piperazin-1-yl) -6-oxohexyl) carbamic acid tert-butyl ester 2c

2b (600mg, 0.87mmol) was dissolved in 5mL of dichloromethane, 5mL of piperidine was added, and the reaction was stirred for 1 hour. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 2c (200mg, yield: 50%).

The third step

N- [ (5R) -5- [ (2R) -2- [ (2R) -2- [ (2R) -2- { [ (tert-butoxy) carbonyl ] amino } -3-phenylpropionamino ] -4-methylpentanylamino ] -6- (4- { [ (4-fluorophenyl) methyl ] carbamoyl } piperazin-1-yl) -6-oxohexyl ] carbamic acid tert-butyl ester 2d

2c (200mg, 0.43mmol), 1k (226mg, 0.43mmol), 2- (7-azobenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (245mg, 0.645mmol) and N, N-diisopropylethylamine (111mg, 0.86mmol) were dissolved in 10mL of N, N-dimethylformamide and reacted with stirring for 12 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 2d (150mg, yield: 36%).

The fourth step

2d (150mg, 0.154mmol) was dissolved in 5mL of dichloromethane, 5mL of a 4M solution of hydrogen chloride in 1, 4-dioxane was added, and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by high performance liquid chromatography to give the title product 2(50mg, yield: 42%).

MS m/z(ESI):773.3[M+1]

¹H NMR(400MHz,CD₃OD)δ7.38-7.31(m,14H),7.24-7.22(m,1H),7.05-7.01(m,2H),5.19-5.13(m,4H),4.85-4.83(m,1H),4.72-4.70(m,1H),4.42-4.40(m,1H),4.34(s,2H),4.07-4.05(m,1H),3.92-3.89(m,2H),3.74-3.46(m,11H),3.25-3.22(m,2H),2.01-1.71(m,9H),1.00-0.95(m,6H).

Example 3

4- ((R) -6-amino-2- ((R) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanoylamino) hexanoyl) -N- (2-methoxyethyl) piperazine-1-carboxamide 3

First step of

4- ((2-methoxyethyl) carbamoyl) piperazine-1-carboxylic acid benzyl ester 3b

Phenyl (2-methoxyethyl) carbamate 3a (2.6g, 13.3mmol, prepared by the method disclosed in patent application "WO 2012002502") and 1d (2.9g, 13.3mmol) were dissolved in 50mL of methanol, and the reaction was stirred at 50 ℃ for 12 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system B to give the title product 3B (2.0g, yield: 50%).

Second step of

N- (2-methoxyethyl) piperazine-1-carboxamide 3c

3b (2.0g, 6.2mmol) was dissolved in 30mL of methanol, palladium on carbon (200mg, 10%) was added, hydrogen gas was substituted three times, and the reaction was stirred for 3 hours. The reaction was filtered and the filtrate was concentrated under reduced pressure to give the crude title product 3c (1g) which was directly used in the next reaction without purification.

The third step

(R) - (9H-fluoren-9-yl) methyl tert-butyl (6- (4- ((2-methoxyethyl) carbamoyl) piperazin-1-yl) -6-oxohexane-1, 5-diyl) dicarbamate 3d

The crude product 3c (500mg, 2.67mmol) and 1g (1.25g, 2.67mmol) were dissolved in 50mL of N, N-dimethylformamide, and 2- (7-azobenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (1.2g, 3.2mmol) and N, N-diisopropylethylamine (0.68mL, 4mmol) were added and reacted with stirring for 12 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 3d (500mg, yield: 30%).

The fourth step

(R) - (5-amino-6- (4- ((2-methoxyethyl) carbamoyl) piperazin-1-yl) -6-oxohexyl) carbamic acid tert-butyl ester 3e

3d (500mg, 0.78mmol) was dissolved in 5mL of dichloromethane, 5mL of piperidine was added, and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give 3e (100mg, yield: 31%).

The fifth step

N- [ (5R) -5- [ (2R) -2- [ (2R) -2- [ (2R) -2- { [ (tert-butoxy) carbonyl ] amino } -3-phenylpropionamino ] -4-methylpentanylamino ] -6- {4- [ (2-methoxyethyl) carbamoyl ] piperazin-1-yl } -6-oxohexyl ] carbamic acid tert-butyl ester 3f

3e (100mg, 0.24mmol), 1k (126mg, 0.24mmol), 2- (7-azobenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (137mg, 0.36mmol) and N, N-diisopropylethylamine (0.1mL, 0.36mmol) were dissolved in 10mL of N, N-dimethylformamide and reacted with stirring for 12 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 3f (80mg, yield: 36.2%).

The sixth step

3f (80mg, 0.086mmol) was dissolved in 2mL of dichloromethane, 2mL of a 4M solution of hydrogen chloride in 1, 4-dioxane was added, and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by high performance liquid chromatography to give the title product 3(10mg, yield: 16%).

MS m/z(ESI):723.4[M+1]

¹HNMR(400MHz,CD₃OD)δ7.38-7.28(m,10H),7.23-7.21(m,1H),5.51(s,2H),4.84-4.82(m,3H),4.75-4.73(m,2H),4.42-4.40(m,1H),4.10-4.08(m,1H),3.71-3.69(m,2H),3.57-3.45(m,6H),3.37(s,3H),3.37-3.36(m,2H),3.25-3.22(m,3H),2.98-2.92(m,5H),1.81-1.61(m,9H),1.34-1.31(m,2H),1.01-0.95(m,6H).

Example 4

(R) -N- ((R) -6-amino-1- (4- (N- (4-fluorobenzyl) aminosulfonyl) piperazin-1-yl) -1-oxohexan-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide 4

First step of

4- (Chlorosulfonyl) piperazine-1-carboxylic acid tert-butyl ester 4b

Sulfonyl chloride (0.52mL, 6.4mmol) was dissolved in 20mL of dichloromethane, 5mL of a solution of piperazine-1-carboxylic acid tert-butyl ester 4a (1g, 5.3mmol) and pyridine (0.63g, 8mmol) in dichloromethane were sequentially added dropwise at 0 ℃, and after completion of the addition, the reaction mixture was warmed to room temperature and stirred for 1 hour. The reaction was poured into 50mL of 1M hydrochloric acid, extracted three times with dichloromethane, the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated under reduced pressure to give the crude title product 4b (1.2g) which was reacted without purification.

Second step of

4- (N- (4-Fluorobenzyl) aminosulfonyl) piperazine-1-carboxylic acid tert-butyl ester 4c

Crude 4b (200mg, 0.7mmol) was dissolved in 20mL acetonitrile and (4-fluorophenyl) methylamine (88mg, 0.7mmol) and cesium carbonate (342mg, 1.05mmol) were added and the reaction stirred at 80 ℃ for 12 h. The reaction solution was cooled to room temperature, filtered, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 4c (50mg, yield: 19.1%).

The third step

N- (4-Fluorobenzyl) piperazine-1-sulfonamide hydrochloride 4d

4c (50mg, 0.134mmol) was dissolved in 1mL of dichloromethane, 1mL of a 4M solution of hydrogen chloride in 1, 4-dioxane was added, and the reaction was stirred for 4 hours. The reaction was concentrated under reduced pressure to give the crude title product 4d (40mg), which was directly used in the next reaction without purification.

The fourth step

(R) - (9H-fluoren-9-yl) methyl tert-butyl (6- (4- (N- (4-fluorobenzyl) sulfamoyl) piperazin-1-yl) -6-oxohexane-1, 5-diyl) dicarbamate 4e

The crude 4d (37mg, 0.134mmol) and 1g (63mg, 0.134mmol) were dissolved in 5mL of N, N-dimethylformamide, and 2- (7-azobenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (76mg, 0.2mmol) and N, N-diisopropylethylamine (34.6mg, 0.268mmol) were added and reacted with stirring for 12 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 4e (70mg, yield: 72.2%).

The fifth step

(R) - (5-amino-6- (4- (N- (4-fluorobenzyl) aminosulfonyl) piperazin-1-yl) -6-oxohexyl) carbamic acid tert-butyl ester 4f

4e (70mg, 0.097mmol) was dissolved in 1mL of dichloromethane, 1mL of piperidine was added, and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give 4f (40mg, yield: 82.5%).

The sixth step

4g of tert-butyl N- [ (5R) -5- [ (2R) -2- [ (2R) -2- [ (2R) -2- { [ (tert-butoxy) carbonyl ] amino } -3-phenylpropionamino ] -4-methylpentanylamino ] -6- (4- { [ (4-fluorophenyl) methyl ] aminosulfonyl } piperazin-1-yl) -6-oxohexyl ] carbamate

4f (40mg, 0.08mmol), 1k (42mg, 0.08mmol), 2- (7-azobenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (45.6mg, 0.12mmol) and N, N-diisopropylethylamine (21mg, 0.16mmol) were dissolved in 10mL of N, N-dimethylformamide and reacted with stirring for 12 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to obtain 4g (30mg, yield: 37%) of the title product.

Seventh step

4g (30mg, 0.03mmol) was dissolved in 1mL of dichloromethane, and 1mL of a 4M solution of hydrogen chloride in 1, 4-dioxane was added thereto, followed by stirring and reaction for 2 hours. The reaction solution was concentrated under reduced pressure to give the title product 4(24mg, yield: 100%).

MS m/z(ESI):809.4[M+1]

¹H NMR(400MHz,CD₃OD)δ8.45-8.41(m,1H),7.41-7.30(m,14H),7.10-7.08(m,2H),4.82-4.80(m,1H),4.74-4.71(m,1H),4.42-4.40(m,1H),4.20-4.18(s,2H),4.18-4.14(m,1H),3.76-3.70(m,4H),3.67-3.37(m,4H),3.21-3.15(m,5H),3.01-2.94(m,6H),1.71-1.50(m,9H),1.02-0.96(m,6H).

Example 5

4- ((R) -6-amino-2- ((R) -2- ((R) -2- ((R) -2-amino-3-phenylpropionylamino) -4-methylpentanoylamino) hexanoyl) -N- (2-fluoroethyl) piperazine-1-carboxamide 5

First step of

4- ((2-fluoroethyl) carbamoyl) piperazine-1-carboxylic acid benzyl ester 5b

Phenyl (2-fluoroethyl) carbamate 5a (387mg, 2.11mmol, prepared by the method disclosed in patent application "US 7253286") and 1d (0.558g, 2.54mmol) were dissolved in 10mL of methanol, heated to 50 ℃ and stirred for 12 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with developer system A to give the title product 5b (0.3g, yield: 97%).

Second step of

N- (2-fluoroethyl) piperazine-1-carboxamide 5c

5b (0.3g, 0.97mmol) was dissolved in 10mL of methanol, palladium on carbon (30mg, 10%) was added, and the reaction mixture was replaced with hydrogen three times, followed by stirring for 4 hours. The reaction was filtered and the filtrate was concentrated under reduced pressure to give the crude title product 5c (0.2g) which was directly used in the next reaction without purification.

The third step

(R) - (9H-fluoren-9-yl) methyl tert-butyl (6- (4- ((2-fluoroethyl) carbamoyl) piperazin-1-yl) -6-oxohexane-1, 5-diyl) dicarbamate 5d

Crude 5c (200mg, 0.97mmol), 1g (455mg, 0.97mmol) was dissolved in 6mL of N, N-dimethylformamide, and 2- (7-azobenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (553mg, 1.46mmol) and triethylamine (0.27mL, 1.94mmol) were added and reacted with stirring for 12 hours. The reaction solution was concentrated under reduced pressure, and ethyl acetate was added to the resulting residue, which was washed with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 5d (600mg, yield: 99%).

The fourth step

(R) - (5-amino-6- (4- ((2-fluoroethyl) carbamoyl) piperazin-1-yl) -6-oxohexyl) carbamic acid tert-butyl ester 5e

5d (200mg, 0.32mmol) was dissolved in 5mL of dichloromethane, 0.5mL of piperidine was added, and the reaction was stirred for 4 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give 5e (110mg, yield: 85%).

The fifth step

N- [ (5R) -5- [ (2R) -2- [ (2R) -2- [ (2R) -2- { [ (tert-butoxy) carbonyl ] amino } -3-phenylpropionylamino ] -4-methylpentanoylamino ] -6- {4- [ (2-fluoroethyl) carbamoyl ] piperazin-1-yl } -6-oxohexyl ] carbamic acid tert-butyl ester 5f

5e (110mg, 0.273mmol), 1k (143mg, 0.73mmol), 2- (7-azobenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (156mg, 0.41mmol) and triethylamine (76uL, 0.546mmol) were dissolved in 5mL of N, N-dimethylformamide and reacted for 4 hours with stirring. The reaction solution was concentrated under reduced pressure, and ethyl acetate was added to the resulting residue, which was washed with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the resulting residue was purified by thin layer chromatography using developer system A to give the title product 5f (145mg, yield: 58%).

The sixth step

5f (145mg, 0.159mmol) was dissolved in 5mL of dichloromethane, 1mL of trifluoroacetic acid was added, and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by high performance liquid chromatography to give the title product 5(57mg, yield: 50%).

MS m/z(ESI):711.3[M+1]

¹H NMR(400MHz,DMSO-d₆):δ8.44(d,2H),8.20(d,3H),7.16-7.37(m,10H),6.01(s,1H),5.11(d,1H),4.32-4.44(m,4H),4.10(d,1H),3.20-3.56(m,14H),2.70(d,2H),1.70-1.80(m,4H),1.48-1.54(m,5H),1.23-1.29(m,2H),1.00(d,3H),0.96(d,3H).

Biological evaluation

The present invention is further described and explained below in conjunction with test examples, which are not intended to limit the scope of the present invention.

Test example 1

1. Purpose of experiment

The aim of this experiment was to test the agonistic action of the compounds of the invention on the kappa opioid receptor (KOR receptor), according to EC₅₀Size compounds were evaluated for in vitro activity.

2. Measurement of KOR Activity

2.1 principle of the experiment

The compounds of the invention can activate the KOR receptor (KOR), thereby reducing the level of intracellular cAMP; the second messenger cAMP enters the cell nucleus to be combined with CRE of DNA, the expression of downstream Luciferase (Luciferase) can be started, the Luciferase can emit fluorescence by reacting with a substrate thereof, and the exciting activity of the compound is reflected by measuring a fluorescence signal.

2.2 Experimental methods

Example compounds agonizing KOR activity affecting changes in downstream cAMP levels was tested by the following method.

2.2.1 Experimental materials and instruments

1) Laboratory apparatus

Name of instrument	Supply company	Model number
			Enzyme-linked immunosorbent assay (ELISA) instrument	PE	Vector3
96-hole round bottom plate	costar	3795
			White 96-well flat bottom plate, transparent bottom	Corning	3903

2) Experimental Material

2.2.2 Experimental procedures

1) Acquisition of HEK293/KOR/CRE monoclonal cell Strain

KOR/pcDNA3.1(+) and CRE/pGL4.29 were transferred into HEK293 cell lines by adding G418 and Hygromycin (Hygromycin) to the medium and HEK293/KOR/CRE monoclonal cell lines were selected in 96-well cell culture plates.

2) EXAMPLES EXPERIMENT OF EXPOCITATION OF COMPOUNDS ON KOR

The HEK293/KOR/CRE monoclonal cell line was cultured in DMEM/high glucose medium (10% FBS,1mg/ml G418, 200ug/ml hygromycin, mixed) and passaged every 3 days. On the day of the experiment, a cell suspension was prepared from fresh cell culture medium, and 20,000 cells/well were plated in a 96-well plate (BD, #356692) and cultured in 5% carbon dioxide at 37 ℃. The next day, compounds were first dissolved in pure DMSO and concentratedThe concentration is 20mM, then DMSO is used for preparing the first concentration of 200nM, the DMSO is used for diluting the first concentration into 8 concentrations in 3 times, 90ul DMSO is added into the blank and control holes; the cells were diluted 20-fold with DMEM/high glucose (SH30243.01B, Hyclone) medium containing 10. mu.M Forskolin. The cell culture plates inoculated on the first day were removed, 10. mu.l of diluted drug or control (0.5% DMSO) was added to each well, mixed by gentle shaking, and incubated at 37 ℃ for 4 hours. In a 96-well cell culture plate, 100ul of luciferase assay solution (Promega, # E6110) was added to each well, allowed to stand at room temperature for 5 minutes, and absorbance was measured using the Victor3.0 luminescence format. EC of the compound was calculated from each concentration of the compound and the corresponding signal value using Graphpad Prism software₅₀The value is obtained. Emax is the maximum effect of a compound to cause changes in cAMP levels.

2.3 test results

Changes in the effect of KOR agonizing on downstream cAMP levels by the Compounds of the invention are determined by the above assay, and the EC measured₅₀The values are shown in Table 1.1.

TABLE 1.1 EC in the agonism of KOR receptor by the Compounds of the invention affecting cAMP levels₅₀

Example numbering	EC₅₀(pM)
		1	24
2	2
		3	13
4	5
		5	9

And (4) conclusion: the compound has obvious agonism on a KOR receptor.

Claims

1. A compound of the general formula (I):

or a pharmaceutically acceptable salt thereof,

wherein:

g is C ═ O or O ═ S ═ O;

R¹is C_1-6Alkyl, wherein said C_1-6Alkyl is optionally selected from halogen, nitro, cyano, hydroxy, C_1-6Alkoxy radical, C_1-6Haloalkoxy, C_1-6Hydroxyalkyl and NR⁶R⁷Is substituted with one or more substituents of (1);

R²selected from halogen, C_1-6Alkoxy, 3-8 membered heterocyclic group and C_6-10Aryl, wherein said 3-to 8-membered heterocyclic group and C_6-10Aryl is optionally substituted with one or more halogens;

R⁶and R⁷Each independently is a hydrogen atom;

and is

n is 1,2, 3 or 4.

2. The compound of formula (I) according to claim 1, which is a compound of formula (II):

or a pharmaceutically acceptable salt thereof,

wherein:

R¹、R²and n is as defined in claim 1.

3. The compound of formula (I) according to claim 1, which is a compound of formula (III):

or a pharmaceutically acceptable salt thereof,

wherein:

G、R²and n is as defined in claim 1.

4. A compound of formula (I) according to claim 1 or 2, wherein R¹Is C_1-6Alkyl, wherein said C_1-6Alkyl being optionally substituted by NR⁶R⁷Substituted by a substituent; r⁶And R⁷Each independently a hydrogen atom.

5. A compound of formula (I) according to claim 1, wherein the compound is selected from:

6. a compound of the general formula (IV):

or a pharmaceutically acceptable salt thereof,

wherein:

R⁶and R⁷Each independently is a hydrogen atom or W;

w is an amino protecting group selected from the group consisting of t-butyloxycarbonyl (Boc), benzyloxycarbonyl, isobutyloxycarbonyl, fluorenylmethyloxycarbonyl (Fmoc), benzoyl, substituted benzoyl, butyryl, acetyl, trifluoroacetyl, phthalimido (Pht), succinimidyl, maleimido, benzyl, allyloxycarbonyl and p-methoxybenzyl;

G、R²and n is as defined in claim 1.

7. The compound of formula (IV) according to claim 6, which is selected from:

8. a process for the preparation of a compound of formula (I) according to claim 1, which process comprises:

wherein:

R⁶and R⁷Each independently is a hydrogen atom or W;

G. n and R²As defined in claim 1.

9. A pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) according to any one of claims 1 to 5, together with one or more pharmaceutically acceptable carriers, diluents or excipients.

10. Use of a compound of formula (I) according to any one of claims 1 to 5 or a pharmaceutical composition according to claim 9 in the preparation of a medicament for the prevention and/or treatment of kappa opioid receptor agonist-mediated disorders.

11. The use according to claim 10 wherein the kappa opioid receptor agonist-mediated related disorder is selected from the group consisting of pain, inflammation, itch, edema, hyponatremia, hypokalemia, ileus, cough and glaucoma.

12. The use according to claim 10 wherein the kappa opioid receptor agonist-mediated related disorder is pain.

13. Use of a compound according to any one of claims 1 to 5 or a pharmaceutical composition according to claim 9 for the manufacture of a medicament for the prevention and/or treatment of pain and pain-related disorders.

14. The use according to claim 13, wherein the pain is selected from neuropathic pain, trunk pain, visceral pain, skin pain, arthritic pain, post-medical treatment pain, ocular pain, otitis pain, breakthrough cancer pain and pain associated with GI disorders.

15. The use according to claim 13, wherein the pain is selected from the group consisting of kidney stone pain, uterine cramps, dysmenorrhea, endometriosis and post-surgical pain.

16. Use of a compound of formula (I) according to any one of claims 1 to 5 or a pharmaceutical composition according to claim 9 in the preparation of a medicament for agonizing the kappa opioid receptor.