RU2784809C2 - Combined product containing dicycloplatin and method for its production and use - Google Patents

Abstract

FIELD: medicine; neurology; therapy.

SUBSTANCE: invention relates to the field of medicine, namely to neurology and therapy; it is intended for the treatment and/or prevention of pain. A method for the treatment and/or prevention of pain includes administration of an effective dose of dicycloplatin to a patient who needs it.

EFFECT: use of the invention allows for prevention and/or elimination of pain for a long period of time with significant efficiency.

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Description

FIELD OF TECHNOLOGY

[1] This invention relates to combined products containing dicycloplatin, the use of combined products for the treatment and/or prevention of pain, inflammation and diseases associated with viral infection, and methods for producing combined products.

BACKGROUND OF THE INVENTION

[2] Since the antitumor effect of cis-dichlorodiamine platinum has been discovered (Rosenberg et al. Nature 1965, 205:698; Nature 1972, 222:385), cisplatin has been widely used as an antitumor drug in clinical medicine. Although these drugs have a therapeutic effect on cancers such as genitourinary tract cancer, nasopharyngeal cancer, cephalosporin-associated cancer, and lung cancer, these drugs are toxic and can cause serious side effects. In the past few decades, the search for highly effective anticancer drugs with low toxicity and a wide spectrum of antitumor activity from platinum analogues has been the subject of research in the field of anticancer drugs.

Structure of Dicycloplatin

[3] Dicycloplatin (DCP) is a co-crystalline compound formed by carboplatin (CBP) and 1,1-cyclobutane dicarboxylic acid (CBDCA) via hydrogen bonding. According to CN1311183A and related academic reports, dicyloplatin has a wide spectrum of antitumor activity, high efficacy, low toxicity, low resistance, low cross-resistance, good penetration, etc. Chinese patent application CN00124864.2 shows that dicycloplatin has a significant effect on head and neck cancer, nasopharyngeal cancer, breast cancer, lung cancer, liver cancer, pancreatic cancer, stomach cancer, intestinal cancer, lymphoma, etc. At the same time, it has a good effect on multiple symptoms such as benign prostatic hyperplasia, prostate inflammation and rheumatoid arthritis (PCT/US2016/028720).

[4] Persistent and breakthrough pain is a major concern for patients with cancer. More than half of all cancer patients will experience severe, uncontrollable pain during the course of their disease, and pain management is a major concern for cancer patients and treating oncologists (List MA, Stracks J, Colangelo L, et al. J. Clin. Oncol ( 2000) 18: 877-884). Although cancer pain is a complex pathological process and a major clinical problem, significant progress has been made in understanding the underlying neurological mechanisms responsible for cancer pain. The symptoms experienced by cancer patients are the result of cellular, tissue and systemic changes that occur during proliferation, invasion and metastasis. A responsive immune system also plays an obvious role in cancer pain. The cancer cell produces mediators that affect other cells in the cancer microenvironment, such as immune cells.

[5] Cancer and immune cells in the area of tumor masses release several neuroimmune mediators that interact with various receptors on peripheral nociceptive nerve endings, promoting abnormal secretions and hyperexcitability. In addition, tumors growing near peripheral nerves can disrupt nerve integrity, causing a neuropathic condition with persistent pain, hyperalgesia, or allodynia. Both of these actions of tumors on the peripheral nerve can lead to central sensitization, which can further increase the efficiency of nociceptive transmission through the dorsal horn and the perception of spontaneous and breakthrough pain (Hamamoto DT, Khasabov SG, Cain DM, Simone DA. J. Neurophysiol (2008 ) 100: 2300-2311, Sabino MA, Luger NM, Mach DB, Rogers SD, Schwei MJ, Mantyh PW Int J Cancer (2003) 104: 550-558).

[6] Platinum-based drugs are rarely used as antiviral drugs. US 8247445 B2 reports the activity of a new platinum complex used in the prevention or treatment of viral and bacterial infections. US 5922689 reports the use of cisplatin derivatives for the treatment of AIDS. Jiajiu Shaw et al. in patent literature (US 6297245 B1) reported that cisplatin was added with folic acid and coenzyme Q10 as a promoter for the treatment of AIDS. Jiajiu Shaw et al. also disclosed that new platinum preparations are used to treat and block new diseases caused by viruses, bacteria and parasites, as well as to treat chronic and inflammatory diseases. Yang Xuqing et al. have disclosed the use of dicycloplatin for antiviral or antibacterial purposes.

[7] However, there remains a need for drugs with good stability and high efficacy in the treatment and/or prevention of pain, inflammation or diseases associated with viral infections.

DISCLOSURE OF THE INVENTION

[8] In a first aspect, the present invention provides a combination product containing dicycloplatin and an acid. In some embodiments, the acid is selected from one or more of the following acids: 1,1-cyclobutanedicarboxylic acid, citric acid, chlorogenic acid, ferulic acid, maleic acid, salicylic acid, folic acid, and gallic acid. In some embodiments, the ratio of dicycloplatin to acids is from 1:0.01 to 1:99. In some embodiments, the combination product optionally contains dicycloplatin and one or more of the following additional components: coenzyme Q10, curcumin, glutathione, oryzanol, citric acid, vitamin C, and anthocyanins. In some embodiments, the combination product may be present in the form of a composition or pharmaceutical composition, or the dicycloplatin, acids, or other ingredients of the combination product may be independently present as separate formulations.

[9] In a second aspect, this invention provides a combination product containing dicycloplatin and one or more of the following additional components: coenzyme Q10, curcumin, glutathione, oryzanol, citric acid, vitamin C and anthocyanins.

[10] In a third aspect, the present invention provides the use of the combination product disclosed earlier herein in the manufacture of a medicament, the medicament being for use in the treatment and/or prevention of pain, inflammation and disease associated with a viral infection. In some embodiments, pain is selected from pain caused by cancer or a cause other than cancer (e.g., hepatitis B, hepatitis C, or rheumatoid arthritis), inflammation is selected from inflammation caused by cancer or a cause other than cancer (e.g., rheumatoid arthritis), the disease associated with a viral infection is selected from diseases associated with infection with hepatitis B virus, hepatitis C virus, Ebola virus or human immunodeficiency virus.

[11] In a fourth aspect, the present invention provides a method for treating and/or preventing pain, inflammation, and disease associated with a viral infection, which comprises administering the combination product described above to patients in need thereof. In some embodiments, pain is selected from pain caused by cancer or a cause other than cancer (e.g., hepatitis B, hepatitis C, or rheumatoid arthritis) and inflammation is selected from inflammation caused by cancer or a cause other than cancer (e.g., rheumatoid arthritis) the disease associated with a viral infection is selected from diseases associated with infection with hepatitis B virus, hepatitis C virus, Ebola virus or human immunodeficiency virus. In some embodiments, the dosage of dicycloplatin in the combined product is 0.01 to 10 mg/kg of the patient's body weight, preferably 0.01 to 5 mg/kg of the patient's body weight. In some embodiments, the combined product can be administered by: orally, by mouth, by inhalation spray, sublingual, rectally, transdermally, through the vaginal mucosa, transmucosally, topically, nasally or enterally, by injection such as intramuscular injection, subcutaneous injection , intramedullary injection and intrathecally, by direct injection into the brain, by in situ injection, subcutaneously, intraperitoneally, intravenously, intraarticularly synovially, intragastrically, intrahepaticly, intralesionly, by intracranial injection, intraperitoneally, by nasal or intraocular injection, or by other methods of drug delivery funds. In some embodiments, the combination product may be in the form of a composition or pharmaceutical composition, or the dicycloplatin, acid, or other ingredients of the combination product may be independently present as separate formulations that may be administered simultaneously, continuously, or at intervals. The time interval includes, without limitation, an integer value from 1 to 24 hours, such as 1 hour, 2 hours, or 3 hours; and an integer value between 1 and 30 days, such as 1 day, 2 days, or 3 days.

[12] In a fifth aspect, the present invention provides a method for producing a combined product, which includes 1) adding dicycloplatinum as an active ingredient or as the sole active ingredient, and 2) optionally adding an acid to form a mixture, wherein the ratio of dicycloplatinum to acid is from 1: 0.01 to 1:99, preferably 1:3 to 1:10, and the acid is selected from one or more of the following: 1,1-cyclobutanedicarboxylic acid, citric acid, chlorogenic acid, ferulic acid, maleic acid, salicylic acid, folic acid and gallic acid, and 3) optionally, the addition of one or more additional ingredients selected from the following: coenzyme Q10, curcumin, glutathione (GSH), oryzanol, citric acid, vitamin C and anthocyanins.

[13] In the fifth aspect, this invention also provides a method for producing a composition containing dicycloplatin or dicycloplatin and 1,1-cyclobutanedicarboxylic acid, including: 1) mixing carboplatin and 1,1-cyclobutanedicarboxylic acid in a molar ratio of from 1:1.05 to 1:99, in a suitable solvent, at room temperature or by heating for an appropriate period of time to obtain a mixture containing dicycloplatin and 1,1-cyclobutanedicarboxylic acid; and optionally 2) directly lyophilizing the mixture, concentrating the mixture, adding an appropriate amount of water to dissolve completely, and then lyophilizing the mixture, or providing the mixture as the desired aqueous solution, thereby obtaining a composition containing dicycloplatin and 1,1-cyclobutanedicarboxylic acid.

[14] In a sixth aspect, the present invention provides the use of dicycloplatin as the sole active ingredient in the manufacture of a medicament, wherein the medicament is for use in the treatment and/or prevention of pain or inflammation, wherein the pain is caused by cancer or another cause (such as a virus hepatitis B, hepatitis C virus, or rheumatoid arthritis), and inflammation is inflammation caused by cancer or a cause other than cancer (eg, rheumatoid arthritis).

[15] In a seventh aspect, the present invention provides the use of the combination product described above for the treatment and/or prevention of pain, inflammation, and disease associated with a viral infection.

[16] In an eighth aspect, the present invention provides a composition containing dicycloplatin as the sole active ingredient and a method for preparing such a mixture, the method comprising adding dicycloplatin as an active ingredient.

[17] In a ninth aspect, the present invention provides the use of dicycloplatin as the sole active ingredient in the manufacture of a medicament, the medicament being for use in the treatment and/or prevention of pain or inflammation. In some embodiments, pain is pain caused by cancer or a cause other than cancer, and inflammation is inflammation caused by cancer or a cause other than cancer (eg, rheumatoid arthritis).

[18] In a tenth aspect, the present invention provides a composition containing dicycloplatin as the sole active ingredient for use in the treatment and/or prevention of pain or inflammation. In some embodiments, the pain is pain caused by cancer or pain caused by a cause other than cancer (e.g., hepatitis B, hepatitis C, or rheumatoid arthritis) and the inflammation is caused by cancer or a cause other than cancer (e.g., rheumatoid arthritis). ).

Figures and description

[19] FIG. 1 shows the effect of dicycloplatin or ribavirin alone on hepatitis C virus (HCV).

[20] FIG. 2 shows the effect of dicycloplatin on hepatitis B virus (HBV).

[21] FIG. 3 shows the effect of dicycloplatin or zidovudine on HIV.

[22] FIG. 4 shows the effect of dicycloplatin on the Ebola virus.

[23] FIG. 5 shows the effect of AA011, VK021 and positive drugs on the virus.

[24] FIG. 6 shows the antiviral effect of the combined composition of MD36792, AA011 (A in Fig. 6) and VK021 (V in Fig. 6).

[25] FIG. 7 shows the analgesic effect of MD36792.

DEFINITION

[26] As used herein, the term "subject" refers to humans and animals, such as dogs, cats, rabbits, chickens, monkeys, etc.

[27] As used herein, the term "dosage" refers to the amount of active ingredient used, for example, milligrams (mg) per kilogram (kg) of a subject's body weight.

[28] Used in this document, the term "ratio" usually refers to the molar ratio. For example, the molar ratio of dicycloplatin to acid is from 1:0.01 to 1:99. The percentage of dicycloplatin refers to the mass percentage.

[29] As used herein, the term "disease associated with a viral infection" refers to a disease caused by a viral infection or a disease accompanying a viral infection. A viral infection includes, without limitation, an infection caused by a hepatitis virus (eg, hepatitis A virus, hepatitis B virus, hepatitis C virus, etc.), Ebola virus, or human immunodeficiency virus.

[30] As used herein, the term "pain" refers to pain caused by a tumor (eg, cancer) or pain caused by another cause (eg, hepatitis B, hepatitis C, or rheumatoid arthritis).

[31] As used herein, the terms "MD36792, AA011, and VK021" refer to dicycloplatin, glutathione, and vitamin C, respectively.

[32] Used in this document, the term "room temperature" means 25±1°C. And, if the experimental temperature is not specified, it refers to room temperature.

[33] As used herein, the term "about" refers to ±10% of the value modified by this term, more preferably ±5%, and most preferably ±2%, so those skilled in the art should clearly define the scope of the term "about" in according to the set value.

DETAILED DESCRIPTION OF THE INVENTION

[34] The authors of this application have found that claimed in this application dicycloplatin as the main active ingredient shows significant efficacy in the treatment and/or prevention of pain, especially pain in patients with cancer. Among cancer patients, especially those with advanced liver cancer, kidney cancer, bone metastases, and bone cancer, the choice is to use dicycloplatin injection or oral formulations as the sole treatment. The inventors found that a certain number of patients with advanced cancer showed no sensitivity to dicycloplatin, and no tumor suppression efficacy was observed. However, dicycloplatin still showed a clear effect on cancer pain relief and even caused pain to disappear. Some patients used dicycloplatin for a certain course of treatment and then stopped treatment, these patients no longer suffered from pain even after they stopped using dicycloplatin.

[35] In the clinical treatment of tumor patients, dicycloplatin was found to have a good effect on tumor-induced edema and tissue inflammation. In many cases, patients with lung cancer with brain metastases have taken dicycloplatin orally or by injection. It was found that in these patients, after using dicycloplatin for one to two weeks, brain tissue edema caused by cancer metastasis was largely eliminated. This shows that dicycloplatin has a good anti-inflammatory effect. In many cases of liver cancer patients with edema of the liver tissue, after infusion of dicycloplatin or oral administration for a certain period of time, the edema of the tissue disappeared or decreased significantly.

[36] In a first aspect, the present invention provides a combination product containing dicycloplatin and acid or consisting of dicycloplatin and acid. In some embodiments, the acid is selected from one or more of the following acids: 1,1-cyclobutanedicarboxylic acid, citric acid, chlorogenic acid, ferulic acid, maleic acid, salicylic acid, folic acid, and gallic acid. In some embodiments, the dicycloplatin to acid ratio is 1:0.01 to 1:99, preferably about 1:1, about 1:1.08, about 1:2, about 1:3, about 1:4, about 1 :5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:20, about 1:30, about 1:40, about 1:50, about 1 :60, about 1:70, about 1:80, about 1:90, and the range of ratios between the ratios includes, without limitation, from about 1:1 to about 1:2, from about 1.08 to about 1:2, from about 1:3 to about 1:10, and so on. In some embodiments, the combination product optionally contains dicycloplatin and one or more of the following additional components: coenzyme Q10, curcumin, glutathione, oryzanol, citric acid, vitamin C, and anthocyanins. In some embodiments, the ratio of dicycloplatin to additional other components is from about 0.1:1 to about 100:1, preferably about 0.1:1, about 0.2:1, about 0.3:1, about 0.4 :1, about 0.43:1, about 0.5:1, about 0.6:1, about 0.7:1, about 0.8:1, about 0.9:1, about 1:1, about 1:1.25, about 1.02:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 20:1, about 30:1, about 40:1, about 50:1, about 60:1, about 70:1, about 80:1, about 90:1, and the range of ratios among the ratios includes, without limitation, from about 0.1:1 to 10:1, from about 0.4:1 to about 8:1, from about 0.43:1 to about 5:1, from about 0 .43:1 to about 1:1.25, from about 0.43:1 to about 1:1.02, and the like. Preferably, the ratio of dicycloplatin to the described additional other components (coenzyme Q10, curcumin, glutathione, oryzanol, citric acid, vitamin C and anthocyanins) is: from about 0.1:1 to 10:1, from about 0.4:1 to about 8 :1 from about 0.43:1 to about 5:1, from about 0.43:1 to about 1:1.25, from about 0.43:1 to about 1:1.02. More preferably, the ratio of dicycloplatin to vitamin C is about 0.43:1, the ratio of dicycloplatin to glutathione is about 1:1.25, the ratio of dicycloplatin to coenzyme Q10 is about 1.02:1. In addition, the weight percentage of dicycloplatin in the compositions is about 5%, about 10%, about 15%, about 16.64%, about 18.29%, about 19.11%, about 19.87%, about 20%, about 20%, about 25%, about 30%, about 35%, about 40%, about 44.46%, about 45%, about 50%, about 55%, about 60%, about 60.5%, about 65% , about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96.2%, about 99%, and is in the range between the corresponding percentages, including, without limitation, from about 5% to 99%, 16.64% to 60.5%, about 18.29% to 44.46%, and about 19.11% to about 35%.

[37] In some embodiments, the combination product may be present in the form of a composition or pharmaceutical composition, or the dicycloplatin, acid, or other ingredients of the combination product may be independently present as separate formulations. In some embodiments, the combination product may be oral formulations such as capsules, tablets, granules, a lyophilized formulation or aqueous injection, an inhaler formulation, or other formulations.

[38] In a second aspect, this invention provides a combination product containing dicycloplatin and one or more of the following additional components: coenzyme Q10, curcumin, glutathione, oryzanol, citrate, vitamin C and anthocyanins. In some embodiments, the combination product optionally contains an acid(s) selected from one or more of the following: 1,1-cyclobutanedicarboxylic acid, citric acid, chlorogenic acid, ferulic acid, maleic acid, salicylic acid, folic acid, and gallic acid. In some embodiments, the ratio of dicycloplatin to acid is from 1:0.01 to 1:99, preferably about 1:1, about 1:1.08, about 1:2, about 1:3, and about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, and the range of ratios among the ratios includes, without limitation, from about 1:1 to about 1:2, from about 1.08 to about 1:2 , from about 1:3 to about 1:10, and so on. In some embodiments, the ratio of dicycloplatin to additional other components is from about 0.1:1 to about 100:1, preferably about 0.1:1, about 0.2:1, about 0.3:1, about 0.4 :1, about 0.43:1, about 0.5:1, about 0.6:1, about 0.7:1, about 0.8:1, about 0.9:1, about 1:1, about 1:1.25, about 1.02:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 20:1, about 30:1, about 40:1, about 50:1, about 60:1, about 70:1, about 80:1, about 90:1, and the range of ratios among the ratios includes, without limitation, from about 0.1:1 to 10:1, from about 0.4:1 to about 8:1, from about 0.43:1 to about 5:1, from about 0 .43:1 to about 1:1.25, from about 0.43:1 to about 1:1.02, and the like. Preferably, the ratio of dicycloplatin to the described additional other components (coenzyme Q10, curcumin, glutathione, oryzanol, citric acid, vitamin C and anthocyanins) is: from about 0.1:1 to 10:1, from about 0.4:1 to about 8 :1 from about 0.43:1 to about 5:1, from about 0.43:1 to about 1:1.25, from about 0.43:1 to about 1:1.02. More preferably, the ratio of dicycloplatin to vitamin C is about 0.43:1, the ratio of dicycloplatin to glutathione is about 1:1.25, the ratio of dicycloplatin to coenzyme Q10 is about 1.02:1. In addition, the weight percentage of dicycloplatin in the compositions is about 5%, about 10%, about 15%, about 16.64%, about 18.29%, about 19.11%, about 19.87%, about 20%, about 20%, about 25% by weight, about 30%, about 35%, about 40%, about 44.46%, about 45%, about 50%, about 55%, about 60%, about 60.5%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96.2%, about 99%, and ranges between their respective percentages, including, but not limited to, about 5% to 99%, 16.64% to 60.5%, about 18.29% to 44.46%, and about 19.11% to about 35%.

[39] In some embodiments, the combination product may be oral formulations such as capsules, tablets, granules, a lyophilized formulation or aqueous injection, an inhaler formulation, or other formulations.

[40] In a third aspect, the present invention provides the use of dicycloplatin, such as the combination product disclosed earlier herein, in the manufacture of a medicament, the medicament being for use in the treatment and/or prevention of pain, inflammation, and disease associated with a viral infection. infection. In some embodiments, pain is selected from pain caused by cancer or a cause other than cancer (e.g., hepatitis B, hepatitis C, or rheumatoid arthritis), inflammation is selected from inflammation caused by cancer or a cause other than cancer (e.g., rheumatoid arthritis) disease associated with viral infection is selected from diseases associated with infection with hepatitis B virus, hepatitis C virus, Ebola virus or human immunodeficiency virus.

[41] In a fourth aspect, the present invention provides a method for treating and/or preventing pain, inflammation, and disease associated with a viral infection, which comprises administering dicycloplatin, such as the combination product described above, to patients in need thereof. In some embodiments, pain is selected from pain caused by cancer or a cause other than cancer (e.g., hepatitis B, hepatitis C, or rheumatoid arthritis) and inflammation is selected from inflammation caused by cancer or a cause other than cancer (e.g., rheumatoid arthritis) the disease associated with a viral infection is selected from diseases associated with infection with hepatitis B virus, hepatitis C virus, Ebola virus or human immunodeficiency virus. In some embodiments, the dosage of dicycloplatin in the combined product is 0.01 to 10 mg/kg of the patient's body weight, preferably 0.01 to 5 mg/kg of the patient's body weight. In some embodiments, the combined product can be administered by: orally, by mouth, by inhalation spray, sublingual, rectally, transdermally, through the vaginal mucosa, transmucosally, topically, nasally or enterally, by injection such as intramuscular injection, subcutaneous injection , intramedullary injection and intrathecal, by direct injection into the brain, by in situ injection, subcutaneous, intraperitoneal, intravenous, intraarticular synovial, intragastric, intrahepatic, intralesional, by intracranial injection, intraperitoneally, by nasal or intraocular injection, or by other methods of drug delivery funds. In some embodiments, the combination product may be in the form of a composition or pharmaceutical composition, or the dicycloplatin, acid, or other ingredients of the combination product may be independently present as separate formulations that may be administered simultaneously, continuously, or at intervals. The time interval includes, without limitation, an integer value from 1 to 24 hours, such as 1 hour, 2 hours, or 3 hours; and an integer value between 1 and 30 days, such as 1 day, 2 days, or 3 days.

[42] In a fifth aspect, the present invention provides a method for producing a combined product, which includes 1) adding dicycloplatinum as an active ingredient or as the only active ingredient, and 2) optionally adding an acid to form a mixture, wherein the ratio of dicycloplatinum to acid is from 1: 0.01 to 1:99, preferably about 1:1, about 1:1.08, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7 , about 1:8, about 1:9, about 1:10, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80 , about 1:90, and the range of ratios among the ratios includes, without limitation, from about 1:1 to about 1:2, from about 1.08 to about 1:2, from about 1:3 to about 1:10, and the acids are selected from one or more of the following, 1,1-cyclobutanedicarboxylic acid, citric acid, chlorogenic acid, ferulic acid, maleic acid, salicylic acid, folic acid, and gallic acid; and 3) optionally, the addition of one or more additional ingredients selected from the following: coenzyme Q10, curcumin, glutathione (GSH), oryzanol, citric acid, vitamin C and anthocyanins, while the ratio of dicycloplatin to additional other components is from about 0, 1:1 to about 100:1, preferably about 0.1:1, about 0.2:1, about 0.3:1, about 0.4:1, about 0.43:1, about 0.5: 1, about 0.6:1, about 0.7:1, about 0.8:1, about 0.9:1, about 1:1, about 1:1.25, about 1.02:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 20:1, about 30:1, about 40:1, about 50:1, about 60:1, about 70:1, about 80:1, about 90:1, and ratios among the ratios include, but are not limited to, from about 0.1: 1 to 10:1, about 0.4:1 to about 8:1, about 0.43:1 to about 5:1, about 0.43:1 to about 1:1.25, from about 0 .43:1 to about 1:1.02, etc.; preferably the ratio of dicycloplatin to other components (coenzyme Q10, curcumin, glutathione, oryzanol, citric acid, vitamin C and anthocyanins) is from about 0.1:1 to 10:1, from about 0.4:1 to about 8:1, about 0.43:1 to about 5:1, about 0.43:1 to about 1:1.25, about 0.43:1 to about 1:1.02; more preferably, the ratio of dicycloplatin to vitamin C is about 0.43:1, the ratio of dicycloplatin to glutathione is about 1:1.25, the ratio of dicycloplatin to coenzyme Q10 is about 1.02:1.

[43] As described herein, an aqueous solution containing dicycloplatin and 1,1-cyclobutanedicarboxylic acid (with a ratio ranging from 1:1.2 to 1:20) is stable at 40°C for seven weeks. An aqueous solution of dicycloplatin with gallic acid, salicylic acid, citric acid, ferulic acid, etc. It is stable at 40°C, but dicycloplatin begins to decompose after two weeks. Preferably, the above acid is 1,1-cyclobutanedicarboxylic acid, and preferably the ratio of dicycloplatin to 1,1-cyclobutanedicarboxylic acid is from 1:1.2 to 1:20.

[44] In some embodiments, this invention provides a method for preparing a composition containing dicycloplatin or dicycloplatin and 1,1-cyclobutanedicarboxylic acid, comprising: 1) mixing carboplatin and 1,1-cyclobutanedicarboxylic acid in a molar ratio of from 1:1.05 to 1:99 (such as 1:1.2, 1:1.5, 1:3, 1:5, 1:10, 1:20 and any ratio between these ratios, including but not limited to 1:1 ,2 to 1:20, or this ratio can be selected according to actual needs, for example, in the proportion described in this document above), in a suitable solvent (preferably water), at room temperature or under heating (preferably 40 ℃ ), for an appropriate period of time, such as 0.5-24 hours (preferably 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 o'clock, 11 o'clock, 12 o'clock, 13 o'clock, 14 o'clock, 15 o'clock, 16 o'clock, 17 o'clock, 18 o'clock, 19 o'clock, 20 o'clock, 21 o'clock, 22 o'clock, 23 hours, 24 hours, or any interval in between) to obtain a mixture containing dicycloplatin and 1,1-cyclobutanedicarboxylic acid; and optionally 2) directly lyophilizing the mixture, concentrating the mixture, adding an appropriate amount of water to dissolve completely, and then lyophilizing the mixture or providing the mixture as the desired aqueous solution, thereby obtaining a composition containing dicycloplatin and 1,1-cyclobutanedicarboxylic acid. Compared to previously reported methods, the present invention has many advantages such as simpler operation, higher yield of dicycloplatin and complete consumption of carboplatin. In addition, the resulting lyophilized mixture consisted of dicycloplatin and 1,1-cyclobutanedicarboxylic acid, and no residue of carboplatin was found.

[45] In the sixth aspect, this invention provides the use of dicycloplatin as an active ingredient or sole active ingredient in the manufacture of a medicinal product, and the medicinal product is intended for use in the treatment and/or prevention of pain or inflammation, while the pain is caused by cancer or another cause ( such as hepatitis B virus, hepatitis C virus, or rheumatoid arthritis), and inflammation is inflammation caused by cancer or a cause other than cancer (eg, rheumatoid arthritis).

[46] In a seventh aspect, the present invention provides a combined product containing dicycloplatin and acid, or consisting of dicycloplatin and acid, for the treatment and/or prevention of pain caused by cancer or other causes other than cancer, such as hepatitis B, hepatitis C or rheumatoid arthritis; inflammation caused by cancer, or another cause not associated with cancer, such as rheumatoid arthritis, and a disease associated with a viral infection, and the disease associated with a viral infection is selected from a disease associated with infection with hepatitis B virus, hepatitis C virus, Ebola or human immunodeficiency virus. In some embodiments, the acid is selected from one or more of the following: 1,1-cyclobutanedicarboxylic acid, citric acid, chlorogenic acid, ferulic acid, maleic acid, salicylic acid, folic acid, and gallic acid. In some embodiments, the composition contains dicycloplatin and one or more of the following other additional components: coenzyme Q10, curcumin, glutathione, oryzanol, citric acid, vitamin C, and anthocyanins.

[47] Alternatively, the combination product contains dicycloplatin and one or more of the following additional components: coenzyme Q10, curcumin, glutathione, oryzanol, citric acid, vitamin C, and anthocyanins. In some embodiments, the compositions optionally contain an acid selected from one or more of the following: 1,1-cyclobutanedicarboxylic acid, citric acid, chlorogenic acid, ferulic acid, maleic acid, salicylic acid, folic acid, and gallic acid.

[48] In some embodiments, the dicycloplatin to acid ratio is from about 1:0.01 to about 1:99, preferably about 1:1, about 1:1.08, about 1:2, about 1:3, about 1 :4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:20, about 1:30, about 1:40, about 1 :50, about 1:60, about 1:70, about 1:80, about 1:90, and the range of ratios between the ratios includes, without limitation, from about 1:1 to about 1:2, from about 1.08 to about 1:2, from about 1:3 to about 1:10, and so on. In some embodiments, the ratio of dicycloplatin to additional other components is from about 0.1:1 to about 100:1, preferably about 0.1:1, about 0.2:1, about 0.3:1, about 0.4 :1, about 0.43:1, about 0.5:1, about 0.6:1, about 0.7:1, about 0.8:1, about 0.9:1, about 1:1, about 1:1.25, about 1.02:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 20:1, about 30:1, about 40:1, about 50:1, about 60:1, about 70:1, about 80:1, about 90:1, and the range of ratios among the ratios includes, without limitation, from about 0.1:1 to 10:1, from about 0.4:1 to about 8:1, from about 0.43:1 to about 5:1, from about 0 .43:1 to about 1:1.25, from about 0.43:1 to about 1:1.02, and the like. Preferably, the ratio of dicycloplatin to the additional other components described (folate, coenzyme Q10, curcumin, glutathione, oryzanol, citric acid, gallic acid, vitamin C and anthocyanins) is: from about 0.1:1 to about 10:1, from about 0, 4:1 to about 8:1 about 0.43:1 to about 5:1, about 0.43:1 to about 1:1.25, about 0.43:1 to about 1:1.02 . More preferably, the ratio of dicycloplatin to vitamin C is about 0.43:1, the ratio of dicycloplatin to glutathione is about 1:1.25, the ratio of dicycloplatin to coenzyme Q10 is about 1.02:1. In some embodiments, the combination product may be present in the form of a composition or pharmaceutical composition, or the dicycloplatin, acid, or other ingredients of the combination product may be independently present as separate formulations.

[49] In an eighth aspect, the present invention provides a composition comprising dicycloplatin as the sole active ingredient, the composition comprising an excipient selected from the following: magnesium stearate, hydroxypropylcellulose, pregelatinized starch, chitosan, β-cyclodextrin, and polyvinylpyrrolidone, and a method for producing such mixtures include the addition of dicycloplatin as the sole active ingredient with the optional excipient described above.

[50] In a ninth aspect, the present invention provides the use of dicycloplatin as an active ingredient or sole active ingredient in the manufacture of a medicament, the medicament being for use in the treatment and/or prevention of pain or inflammation. In some embodiments, the pain is pain caused by cancer or pain caused by a cause other than cancer (e.g., hepatitis B, hepatitis C, or rheumatoid arthritis), and the inflammation is inflammation caused by cancer or inflammation caused by a cause other than cancer. from cancer (for example, rheumatoid arthritis).

[51] In a tenth aspect, the present invention provides compositions containing dicycloplatin as the active ingredient or the sole active ingredient, for use in the treatment and/or prevention of pain or inflammation. In some embodiments, the pain is pain caused by cancer or pain caused by a cause other than cancer (e.g., hepatitis B, hepatitis C, or rheumatoid arthritis) and the inflammation is caused by cancer or a cause other than cancer (e.g., rheumatoid arthritis). ).

[52] In other aspects, the present invention provides a method for treating and/or preventing pain, comprising administering an effective dose of dicycloplatin to a patient in need thereof. In some embodiments, the patient is a patient with cancer, preferably a patient suffering from liver cancer, kidney cancer, bone metastases, brain metastases, or bone cancer, more preferably a patient with advanced cancer, or a patient who has already developed resistance to an anticancer drug. a platinum-based drug, most preferably in a patient with advanced liver cancer. In addition, the effective dose of dicycloplatin may be lower than the effective anticancer dose of dicycloplatin in a cancer patient, or in some embodiments, the effective dose of dicycloplatin may be equal to the effective dose of the anticancer agent used by the patient with cancer. For example, dicycloplatin can be used as a single drug for the treatment of cancer and pain. In some embodiments, dicycloplatin may be used as the sole active agent for the treatment and/or prevention of pain. In some embodiments, the implementation of the method of treatment further includes the use of one or more other active substances that have an analgesic effect. In some embodiments, the method further comprises administering one or more anticancer actives, wherein the anticancer active is preferably a non-platinum based anticancer drug. In some embodiments, the dicycloplatin is present in the form of a pharmaceutical composition. In addition, dicycloplatin is contained in the combined product as defined herein above.

[53] In other aspects of the invention, the effective dose of dicycloplatin for the treatment and/or prevention of pain may be lower than the effective anticancer dose, and dicycloplatin has an analgesic effect in cancer patients who are already resistant to the antitumor activity of platinum drugs. Therefore, the methods or uses of dicycloplatin for the treatment and/or prevention of pain are not limited to patients with cancer, and dicycloplatin has demonstrated a significant analgesic effect comparable to positive drug control rotunidine in the animal pain model experiments shown herein. As shown herein, dicycloplatin can have an analgesic effect on pain caused by cancer or non-cancer, including, without limitation, hepatitis B, hepatitis C, and rheumatoid arthritis. In some embodiments, in the above method and use for the treatment and/or prevention of pain, the patient in need thereof is not a cancer patient. In some embodiments, in the above method and use for the treatment and/or prevention of pain, the patient in need thereof is a cancer patient who has developed resistance to a platinum-based anticancer drug.

[54] In other aspects, the invention relates to a method for treating and/or preventing tissue edema (e.g., liver edema, cerebral edema, etc.) or inflammation, the method comprising administering an effective dose of dicycloplatin to a patient in need thereof. In some embodiments, the patient is a patient with cancer, preferably a patient with liver cancer, kidney cancer, bone metastases, brain metastases, or bone cancer, more preferably a patient with advanced cancer, most preferably a patient with advanced liver cancer, or a patient who is resistant to a platinum-based anti-cancer drug. In some embodiments, an effective dose of dicycloplatin may be lower than an effective anticancer dose of dicycloplatin in a cancer patient, or in some embodiments, an effective dose of dicycloplatin may be equal to an effective dose of an anticancer agent used by a patient with cancer. In some embodiments, the method further comprises administering to the patient one or more anti-cancer actives, wherein the anti-cancer actives are preferably a non-platinum-based anti-cancer drug. In some embodiments, the patient is not a cancer patient. In some embodiments, the dicycloplatin is present in the form of a pharmaceutical composition. In addition, dicycloplatin is contained in the combined product as defined herein above.

[55] In other aspects, the present invention provides a method for treating and/or preventing a disease associated with a viral infection, such as a method for lowering a viral index, comprising administering an effective dose of a pharmaceutical composition to a patient in need thereof, wherein the pharmaceutical composition comprises dicycloplatin and glutathione. In some embodiments, the viral infection is an infection with hepatitis B virus, hepatitis C virus, Ebola virus, and/or human immunodeficiency virus. In some embodiments, the viral infection is a hepatitis B virus infection. In some embodiments, the viral infection is a hepatitis C virus infection. In some embodiments, the viral infection is an Ebola virus infection. In some embodiments, the viral infection is a human immunodeficiency virus infection. In some embodiments, the method includes administering to patients an effective dose of the pharmaceutical compositions 1 to 3 times per day. In some embodiments, the method includes administering to the patient an effective dose of the pharmaceutical composition for at least 7 days, preferably 1 month to 1 year, such as 1, 2, 3, 4, 5, 6, 7. 8, 9, 10 , 11, 12 months, preferably 1 to 6 months, eg 1, 2, 3, 4, 5, 6 months. In some embodiments, the patient is not being treated with any other antiviral drug and/or has not received any other antiviral drug treatment. In some embodiments, the method also includes administering other antiviral drugs to the patient. In some embodiments, the pharmaceutical composition optionally contains dicycloplatin and one or more of the following other additional components: coenzyme Q10, curcumin, oryzanol, citric acid, vitamin C, and anthocyanins. Preferably, the ratio of dicycloplatin to glutathione is from about 0.1:1 to about 100:1, preferably about 0.1:1, about 0.2:1, about 0.3:1, about 0.4:1, about 0.43:1, about 0.5:1, about 0.6:1, about 0.7:1, about 0.8:1, about 0.9:1, about 1:1, about 1:1 .25, about 1.02:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 20:1, about 30:1, about 40:1, about 50:1, about 60:1, about 70:1, about 80:1, about 90:1, and the range of ratios among the ratios includes, without limitation, from about 0.1:1 to 10:1, from about 0.4:1 to about 8:1, from about 0.43:1 to about 5:1, from about 0.43:1 to about 1:1.25, from about 0.43:1 to about 1:1.02, and the like. The ratio of dicycloplatin to glutathione is about 0.1:1 to 10:1, about 0.4:1 to about 8:1, about 0.43:1 to about 5:1, about 0.43:1 to about 1:1.25, from about 0.43:1 to about 1:1.02. More preferably, the ratio of dicycloplatin to glutathione is about 1:1.25. Preferably, dicycloplatin and glutathione are present in the same pharmaceutical composition in the treatment of diseases associated with viral infections. However, in some embodiments, dicycloplatin and glutathione may be independently present as separate formulations that may be administered simultaneously, continuously, or at regular intervals.

[56] In some embodiments, in the above method of treating and/or preventing pain, inflammation, and diseases associated with viral infections, dicycloplatin or pharmaceutical compositions may be present in the form of oral formulations such as capsules, tablets, granules, or frozen dry formulations, or aqueous injections, or formulations for inhalers, or other formulations. In some embodiments, in the above described method for the treatment and/or prevention of pain, inflammation, and diseases associated with viral infections, the dicycloplatin or pharmaceutical composition can be administered as follows: orally, in the mouth, by inhalation spray, sublingually, rectally, transdermally, through the vaginal mucosa , transmucosally, by topical administration, by nasal or intestinal administration, by injection administration such as intramuscular, subcutaneous, intramedullary and intrathecal, by direct injection into the brain, by orthotopic administration, subcutaneous, intraperitoneal, intravenous, intraarticular synovial, intragastric, intrahepatic , intralesional, via intracranial administration, intraperitoneally, via nasal or intraocular injection, or via other drug delivery methods. Preferably, the dicycloplatin or pharmaceutical composition may be administered orally or by injection.

DETAILED DESCRIPTION

[57] The present invention is further described below with reference to the figures and specific embodiments. It should be understood that the following examples are used only to illustrate the present invention and are not used to limit the scope covered by the claims of the present invention.

[58] Dicycloplatin, mentioned in the examples, can be purchased commercially or obtained by the methods described in this document. Other compounds or acids and other components described herein are well known to those skilled in the art, may be purchased commercially, or obtained using known synthetic methods.

DETECTION METHODS

[59] The detection methods used in the examples in this article are as follows:

(1) X-ray powder diffractometry (XRPD)

[60] Analytical instrument: Panalytical Empyrean.

[61] X-ray powder diffraction was performed by mounting a sample of crystalline material on a silicon (Si) low background single crystal holder spreading the sample into a thin layer using a microscope slide. Position 2i was calibrated to the Panalytical 640 Si powder standard. The sample was irradiated with X-rays generated by a long fine focusing copper tube operating at 45 kV and 40 mA with a wavelength of Kα1 = 1.540589 angstroms and Kα2 = 1.544426 angstroms (intensity ratio Kα1/Kα2 is 0.50). A collimated X-ray source was passed through a programmed divergence slit set to 10 mm and reflected radiation was directed through a 5.5 mm anti-scatter slit. The sample was exposed for 16.3 seconds in 0.013° 2-theta increments (continuous scan mode) from 3 degrees to 40 degrees 2-theta in theta-theta mode. The running time was 3 minutes 57 seconds. The instrument was equipped with an RTMS detector (XʹCelerator). Management and data collection was performed using a Dell Optiplex 780 XP running data collection software.

[62] As is known, the following factors can affect the relative intensity of the peaks, for example, grains larger than 30 microns and non-unitary aspect ratios, which can affect the analysis of samples. In addition, the position of the reflections can be affected by the exact height at which the sample is in the diffractometer and the zero calibration of the diffractometer. The flatness of the sample surface may also have a small effect. Therefore, the presented diffraction pattern data should not be taken as absolute values.

(2) Differential Scanning Calorimetry (DSC)

[63] As a thermal analysis method, DSC is used to measure the amount of heat required to heat samples and also measure a reference value as a function of temperature. The general procedure for measuring DSC is known. The specific tools and conditions used in the examples described are as follows:

Analytical instrument: TA Instruments Q2000 DSC.

Heating rate: 10°C per minute.

Purge gas: nitrogen

Thermogravimetric analysis (TGA)

[64] TGA is used to measure the change in the physical and chemical properties of a sample with elevated temperature, typically at a constant heating rate or over time (at constant temperature and/or constant mass loss). The general procedure for measuring TGA is known. The specific tools and conditions used in the examples described are as follows:

Analytical instrument: TA Instruments Q5000 TGA.

Heating rate: 10°C per minute.

Purge gas: nitrogen.

Example 1 Preparation of Dicycloplatin Compositions

[65] 1) 5.0 g of carboplatin (13.47 mmol) and 2.232 g of 1,1-cyclobutanedicarboxylic acid (15.50 mmol) were added to a 20 ml glass flask.

2) 15 ml of deionized water was added to the above glass vial and stirred at room temperature to form a homogeneous suspension, keeping the vial away from the light source.

3) Cooled to 20°C, the resulting mixture was transferred into a reaction flask with a volume of 500 ml and washed with 50 ml of deionized water.

4) Added another 175 ml of deionized water, mixed well and kept at 20°C for 2 hours to form a homogeneous solution.

5) The mixture was filtered. The filtrate was lyophilized and dried in vacuo.

6) 7.21 g of a mixture of dicycloplatin and 1,1-cyclobutanedicarboxylic acid were obtained.

7) It was established by measurement that the dicycloplatin amounted to 96.20% of the mass. the ratio of DCP/1,1-cyclobutanedicarboxylic acid was 1:0.15.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and ¹ H NMR as described above.

Example 2 Preparation of Dicycloplatin Compositions

[66] 1) 5.0 g of carboplatin (13.47 mmol) and 22.32 g of 1,1-cyclobutanedicarboxylic acid (155.0 mmol) were added to a 200 ml glass flask.

2) 125 ml of deionized water was added to the above glass vial and stirred at room temperature to form a homogeneous suspension, keeping the vial away from the light source.

3) Cooled to 20°C, the resulting mixture was transferred to a 500 ml reaction flask and washed with 150 ml of deionized water.

4) Added another 2000 ml of deionized water, mixed well and kept at 20°C for 2 hours to form a homogeneous solution.

5) The mixture was filtered. The filtrate was lyophilized and dried in vacuo.

6) 72.1 g of a mixture of dicycloplatin and 1,1-cyclobutanedicarboxylic acid were obtained.

7) It was determined by measurement that dicycloplatin was 96.22% by weight, the ratio DCP/1,1-cyclobutanedicarboxylic acid was 1:10.15.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and ¹ H NMR as described above.

Example 3 Preparation of a combined product containing 1,1-cyclobutanedicarboxylic acid, citric acid and dicycloplatin

[67] 1) 5.0 g of carboplatin (13.47 mmol) and 3.879 g of 1,1-cyclobutanedicarboxylic acid (26.94 mmol) were added to a 50 ml glass flask.

2) 25 ml of deionized water was added to the above glass vial and stirred at room temperature to form a homogeneous suspension, keeping the vial away from the light source.

3) Cooled to 20°C, the resulting mixture was transferred into a reaction flask with a volume of 500 ml and washed with 50 ml of deionized water.

4) Added another 225 ml of deionized water, mixed well and kept at 20°C for 2 hours to form a homogeneous solution.

5) 2.588 g (13.47 mmol) of citric acid was added to the above solution and stirred until a homogeneous solution was formed.

6) The mixture was filtered. The filtrate was lyophilized and dried in vacuo.

7) 11.462 g of a mixture of dicycloplatin and 1,1-cyclobutanedicarboxylic acid were obtained.

8) It was established by measurement that dicycloplatin was 60.50% by weight. ratio DCP/1,1-cyclobutanedicarboxylic acid/citric acid: 1:1:1.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and ¹ H NMR as described above.

Example 4 Preparation of a combined product containing 1,1-cyclobutanedicarboxylic acid, citric acid, GSH and dicycloplatinum

[68] 1) 5.0 g of carboplatin (13.47 mmol) and 3.879 g of 1,1-cyclobutanedicarboxylic acid (26.94 mmol) were added to a 50 ml glass flask.

2) 25 ml of deionized water was added to the above glass vial and stirred at room temperature to form a homogeneous suspension, keeping the vial away from the light source.

3) Cooled to 20°C, the resulting mixture was transferred into a reaction flask with a volume of 500 ml and washed with 50 ml of deionized water.

4) Added another 225 ml of deionized water, mixed well and kept at 20°C for 2 hours to form a homogeneous solution.

5) 2.588 g (13.47 mmol) of citric acid and 4.135 g of glutathione (GSH, 13.47 mmol) were added to the above solution and stirred until a homogeneous solution was formed.

6) The mixture was filtered. The filtrate was lyophilized and dried in vacuo.

7) 15.60 g of a mixture of dicycloplatin, glutathione and 1,1-cyclobutanedicarboxylic acid were obtained.

8) It was established by measurement that the dicycloplatin amounted to 44.46% of the mass. ratio DCP/1,1-cyclobutanedicarboxylic acid/citric acid/GSH: 1:1:1:1.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and ¹ H NMR as described above.

Example 5 Preparation of a combined product containing 1,1-cyclobutanedicarboxylic acid, chlorogenic acid, glutathione, vitamin C and dicycloplatinum

[69] 1) 5.15 g dicycloplatinum (13.47 mmol), 2.232 g 1,1-cyclobutanedicarboxylic acid (15.50 mmol), 4.77 g chlorogenic acid (13.47 mmol), 4.135 g glutathione (GSH , 13.47 mmol)), 11.862 g vitamin C was mixed well.

2) 28.15 g of a dicycloplatin-containing combined product was then obtained.

7) The content of dicycloplatin was up to 18.29% of the mass.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and ¹ H NMR as described above.

Example 6 Preparation of a combined product containing chlorogenic acid, glutathione, vitamin C and dicycloplatin

[70] 1) 5.15 g dicycloplatinum (13.47 mmol), 4.77 g chlorogenic acid (13.47 mmol), 4.135 g glutathione (GSH, 13.47 mmol) and 11.862 g vitamin C were thoroughly mixed.

2) Received 25.92 g of combined products with dicycloplatin.

7) The content of dicycloplatinum was 19.87%.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and ¹ H NMR as described above.

Example 7 Preparation of a combined product containing glutathione, vitamin C and dicycloplatin

[71] 1) 5.15 g of dicycloplatin (13.47 mmol), 4.135 g of glutathione (GSH, 13.47 mmol) and 11.862 g of vitamin C were thoroughly mixed.

2) 21.147 g of dicycloplatin-containing combined product was obtained.

7) The mass of dicycloplatinum was 24.35%.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and 1H NMR as described above. Example 8 Preparation of a combined product containing folic acid, chlorogenic acid, glutathione, vitamin C and dicycloplatin

[72] 1) 5.15 g dicycloplatinum (13.47 mmol), 1.03 g folic acid, 4.77 g chlorogenic acid (13.47 mmol), 4.135 g glutathione (GSH, 13.47 mmol) and 11.862 g vitamin C was thoroughly mixed.

2) 26.95 g of a dicycloplatin-containing combined product was obtained.

7) The mass of dicycloplatinum was 19.11%.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and ¹ H NMR as described above.

Example 9 Preparation of a combined product containing coenzyme Q10, chlorogenic acid, glutathione, vitamin C and dicycloplatin

[73] 1) 5.15 g dicycloplatin (13.47 mmol), 5.03 g coenzyme Q10, 4.77 g chlorogenic acid (13.47 mmol), 4.135 g glutathione (GSH, 13.47 mmol) and 11.862 g vitamin C was thoroughly mixed.

2) 30.95 g of a dicycloplatin-containing combined product was obtained.

7) The mass of dicycloplatinum was 16.64%.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and ¹ H NMR as described above.

Example 10 Preparation of a combined product containing coenzyme Q10, citric acid, chlorogenic acid, glutathione, vitamin C and dicycloplatin

[74] 1) 5.15 g dicycloplatin (13.47 mmol), 5.03 g coenzyme Q10, 4.77 g citric acid, 4.135 g glutathione (GSH, 13.47 mmol) and 11.862 g vitamin C were thoroughly mixed.

2) 30.95 g of a dicycloplatin-containing combined product was obtained.

7) The mass of dicycloplatinum was 16.64%.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and ¹ H NMR as described above.

Example 11 Preparation of a combined product containing oryzanol, citric acid, glutathione, vitamin C and dicycloplatin

[75] 1) 5.15 g of bicyclic platinum (13.47 mmol), 5.03 g of oryzanol, 4.77 g of citric acid, 4.135 g of glutathione (GSH, 13.47 mmol) and 11.862 g of vitamin C were thoroughly mixed.

2) 30.95 g of a dicycloplatin-containing combined product was obtained.

7) The mass of dicycloplatinum was 16.64%.

The product was analyzed by XRPD (X-ray powder diffraction), DSC (differential scanning calorimetry) and ¹ H NMR as described above.

Example 12 In Vitro Activity Study of MD36792 (Dicycloplatin) against Hepatitis C Virus (HCV)

[76] (I) Experimental materials:

1. Compounds: MD36792 (dicycloplatin) and ribavirin (commercially available).

2. Cell: Huh7.5.1 cells.

3. Virus: Hepatitis C virus (HCV type 2a, virus strain JFH1-Luc).

4. Experimental reagents: DMEM medium, alamarBlue® kit (Invitrogen), Luciferase Assay System kit (Promega) and other experimental reagents.

[77] (II) Experimental methods

1. Cytotoxicity test MD36792

[78] Experimental principle: The experiment uses the alamarBlue® kit (Invitrogen) to determine the toxic effect of drugs on cells. alamarBlue® is a redox indicator that changes in absorbance and fluorescent signals based on cellular metabolic activity. alamarBlue® dissolves easily in water. Once its oxidized form enters cells, it is followed by reduction by mitochondrial enzymes to induce measurable fluorescence and color change. It is used for quantitative analysis of cell viability and proliferation as well as in vitro cytotoxicity studies. This assay is based on the ability of cells with metabolic activity to convert reagents into fluorescent and colorimetric indicators. Damaged and inactive cells have lower natural metabolic activity and lower corresponding signals, so the strength of the fluorescent signal can reflect the level of cell activity.

[79] Method steps: Huh7.5.1 cells were seeded in a 96-well cell culture plate, and attached after use. DMSO was used to continuously dilute the drug from 200-fold initial concentration to a 3-fold gradient by 6 gradients, and then complete DMEM was used to dilute the drug-containing culture solution, and each well was tested in duplicate. After 72 hours of incubation, the culture supernatant was discarded and medium containing alamarBlue® was added. After incubation at 37° C. for 4 hours, fluorescence readings were detected at 570 nm and 595 nm.
Calculation formula: cell viability (%) = (sample well-blank control) / (control cell-blank control) * 100%

2. Hepatitis C virus (HCV) experiment MD36792

[80] Experimental principle: After the HCV strain JFH1-Luc containing the Rluc reporter gene infected Huh7.5.1 cells, Rluc was expressed as a replicated virus, and the expression level of the Rluc reporter gene could reflect the replication level of the HCV virus.

Experimental steps: (1) Cell seeding: Huh7.5.1 cells were seeded in 96-well cell culture plates, and the cells were attached to the wall for later use. (2) Viral infection: The drug was serially diluted from 2 times as the initial concentration and 3 times from gradient to 9 gradients in the culture medium. Each concentration was in 2 replicate wells. 100 μl of the drug-containing culture solution was added to the cell wells and then infected with 100 μl of the JFH1-luc virus culture medium. (3) Detection: cultured for 72 hours after virus infection. Rluc readings were determined using the Renilla Luciferase Assay.

[81] In the experiment, a ribavirin-positive drug control group, a viral control group, and a cellular control group were selected.

[82] Calculation formula: inhibition ratio (%) = 100% - (drug group - cell control group) / (viral control group - cell control group) * 100%

[83] Experimental results: As shown in FIG. 1, MD36792 has an EC ₅₀ (50% inhibition) of HCV of 121.1 μM, so the SI selection index (SI = CC ₅₀ / EC ₅₀ ) of MD36792 is 1.54. Because HCV replication in this model requires a high cellular status, when a drug has a cytotoxic effect, it will seriously affect viral replication. Therefore, we believe that the inhibitory effect of 333 μM and 111 μM against HCV is due to toxic effects. Therefore, MD36792 has no inhibitory effect on HCV. The control drug ribavirin has a significant inhibitory effect on HCV.

Example 13 In Vitro Activity Study of MD36792 (Dicycloplatin) against Hepatitis B Virus (HBV)

[84] (I) Experimental materials:

1. Connection: MD36792.

2. Cell: HepAD38 cells.

3. Virus: hepatitis B virus (HBV).

4. Experimental reagents: DMEM/F12 medium, CellTiter-Glo detection kit (Promega), HBeAg antigen detection kit (Kehua), HBsAg antigen detection kit (Kehua) and other experimental reagents.

[85] (II) Experimental methods

1. Cytotoxicity test MD36792

[86] Experimental principle: CellTiter-Glo kit (Promega) was used to detect the toxic effects of drugs on cells. The CellTiter-Glo kit determines the number of living cells in culture by quantifying ATP. The respiration of metabolically active cells and other vital activities can produce ATP. The stable type of luminescence produced by luciferase is used in the Signaling kit, while luciferase requires ATP in the light emission process. An equal volume of CellTiter-Glo reagent was added to the cell culture medium to measure the amount of luminescence. The light signal is proportional to the amount of ATP in the system, and ATP is positively correlated with the number of living cells. Therefore, the value of the light signal may reflect the number of living cells.

[87] Method steps: HepAD38 cells were seeded in a 96-well cell culture plate, and attached to the wall for later use. The drug was serially diluted from 1 mM to 9 gradients in 9 gradients with 2 replicates per gradient. The drug was added to the cells and cultured in a CO ₂ incubator at 37°C. After 3 days of drug addition and culture, the drug-induced cytopathic effect (CPE) was observed under a light microscope, and CellTiter-Glo was added to determine cell viability. Cell toxicity of a drug is expressed as cell activity.

[88] Calculation formula: cell activity (%) = (drug group-blank control) / (control cell-blank control) * 100%

2. Hepatitis B virus (HBV) experiment MD36792

[89] Experimental principle: HBV replication in the HepAD38 cell line can be regulated by tetracycline. After removing dox, the integrated HBV genome can be transcribed to form pgRNA (pregenomic RNA), and pgRNA continues reverse transcription into ocDNA (open circular DNA). The synthesized ocDNA will re-enter the nucleus to form cncDNA (covalently continuous circular DNA). HBeAg can only be expressed from cncDNA, and HBeAg expression can reflect the amount of cncDNA synthesis. Therefore, we use ELISA to determine the content of HBeAg and determine the inhibitory effect of the drug on HBV. In this cell, HBsAg can be continuously transcribed and translated from integrated HBV DNA. We also tested the HBsAg content to determine the effect of the drug on HBV replication.

[90] Experimental steps: (1) Cell seeding: HepAD38 cells with Dox removed for 3 days were seeded in 96-well plates and cultured in a CO ₂ incubator at 37°C for 24 hours. (2) Dosing: MD36792 was diluted with complete DMEM/F12 (10% PBS) from 333 µM, diluted with 9 gradients 3 times in a row and 2 x 2 replicate wells per gradient. The diluted preparation was added to the cells and cultured in a CO ₂ incubator at 37°C for 72 hours. (3) Detection: Take the supernatant and use the HBeAg antigen detection kit and the HBsAg antigen detection kit to detect HBeAg and HBsAg expression respectively. Calculation formula: degree of antigen inhibition (%) = 100% - (drug group - blank control) / (cell control group - blank control) * 100%

[91] Results of the experiment. As shown in FIG. 2, MD36792 had no inhibitory effect on the expression of HBeAg (e antigen) and HBsAg (surface antigen) of HBV.

Example 14 In Vitro Activity Study of MD36792 (Dicycloplatin) against Human Immunodeficiency Virus (HIV)

[92] (I) Experimental materials:
1 Compounds: MD36792 and zidovudine (AZT) (commercially available).
2. Cell: MT4 cells.
3. Virus: human immunodeficiency virus (HIV).
4. Experimental reagents: RPMI-1640 medium, CellTiter-Glo detection kit (Promega) and other experimental reagents.

[93] (II) Experimental methods

1. Cytotoxicity test MD36792

[94] Experimental principle: CellTiter-Glo kit (Promega) was used to detect the toxic effect of drugs on cells. The CellTiter-Glo kit determines the number of living cells in culture by quantifying ATP. The respiration of metabolically active cells and other vital activities can produce ATP. The stable type of luminescence produced by luciferase is used in the Signaling kit, while luciferase requires ATP in the light emission process. An equal volume of CellTiter-Glo reagent was added to the cell culture medium to measure the amount of luminescence. The light signal is proportional to the amount of ATP in the system, and ATP is positively correlated with the number of living cells. Therefore, the value of the light signal may reflect the number of living cells.

[95] Method steps: MT4 cells were seeded in a 96-well cell culture plate, and attached to the wall for later use. The drug was serially diluted from 1 mM to 9 gradients in 9 gradients with 2 replicates per gradient. The drug was added to the cells and cultured in a CO ₂ incubator at 37°C. The drug-induced cytopathic effect (CPE) was observed under a light microscope and CellTiter-Glo was added to determine cell viability. Cell toxicity of a drug is expressed as cell activity.

[96] Calculation formula: cell activity (%) = (drug group-blank control) / (control cell-blank control) * 100

2. MD36792 experiments on human immunodeficiency virus (HIV)

[97] Experimental principle: MT-4 cell protection experiments were used to detect antiviral compounds.

[98] Experimental steps: A 3-fold diluted drug was added to a 96-well plate in which LAI virus and MT-4 cells were incubated, and they (cells) were inoculated into the well plate. After cultivation, when the virus infected the control wells, there was a significant CPE. The CellTiter-Glo reagent was added and mixed, then the chemiluminescence signal was detected.

[99] Cellular control (control of non-infected cells), viral control and positive drug control (zidovudine, AZT) were isolated in the experiment.

[100] Inhibition ratio (%) = (drug group-viral control) / (cell control-viral control) * 100% Experimental results: as shown in FIG. 3, MD36792 has no inhibitory effect on HIV infection, while the control drug zidovudine has a significant inhibitory effect on HIV.

Example 15 In vitro study of the activity of MD36792 (bicyclic platinum) against Ebola virus

[101] (I) Experimental materials

1. Connection: MD36792.

2. Cells: MDCK cells.

3. Virus: Ebola virus.

4. Experimental reagents: DMEM medium, alamarBlue® kit (Invitrogen), Luciferase Assay System kit (Promega) and other experimental reagents.

[102] (II) Experimental methods

1. Cytotoxicity test MD36792

[103] Experimental principle: The experiment uses the alamarBlue® kit (Invitrogen) to determine the toxic effect of drugs on cells. alamarBlue® is a redox indicator that changes in absorbance and fluorescent signals based on cellular metabolic activity. alamarBlue® dissolves easily in water. Once its oxidized form enters cells, it is followed by reduction by mitochondrial enzymes to induce measurable fluorescence and color change. It is used for quantitative analysis of cell viability and proliferation as well as in vitro cytotoxicity studies. This assay is based on the ability of cells with metabolic activity to convert reagents into fluorescent and colorimetric indicators. Damaged and inactive cells have lower natural metabolic activity and lower corresponding signals, so the strength of the fluorescent signal can reflect the level of cell activity.

[104] Experimental procedure. MDCK cells were seeded in a 96-well cell culture plate and attached to wait. The drug was serially diluted from 1 mM to 9 gradients in 9 gradients with 2 replicates per gradient. The drug was added to the cells and cultured in a CO ₂ incubator at 37°C. After 2 days of dosing and incubation, the drug-induced cytopathic effect (CPE) was observed under a light microscope. Then alamarBlue® medium was added and incubated at 37°C for 4 hours, then fluorescence readings were determined at 570 nm and 595 nm. Cell toxicity of a drug is expressed as cell activity.

[105] Calculation formula: cell activity (%) = (drug group-blank control) / (control cell-blank control) * 100%

2. Experiment: MD36792 against the Ebola virus

[106] The principle of the experiment: after the Ebola virus containing the Rluc reporter gene infects cells, Rluc is expressed when the virus replicates. The expression level of the Rluc reporter gene may reflect the level of adenovirus infection.

[107] Method steps: (1) Cell seeding: MDCK cells were seeded in a 96-well cell culture plate and used after attachment. (2) Dose treatment: The drug was gradually diluted 3 times into 9 groups, according to drug concentration, with maintenance medium (2% PBS), and combined into a culture solution containing 2 times the drug dose with 2 replicates per gradient . After the cells had grown into a monolayer, 100 μl of 2x DMEM-containing culture medium was added, followed by 100 μl of DMEM-diluted Ebola virus and incubated at 37°C. (3) Detection: After culturing for 48 hours after virus infection, Rluc readings were determined using a luciferase assay kit.

[108] In the experiment, a viral control group and a cellular control group were isolated.

[109] Calculation formula: inhibition ratio (%) = 100% - (drug group-cell control group) / (viral control group-cell control group) * 100%

[110] Results of the experiment. As shown in FIG. 4, MD36792 has no inhibitory effect on Ebola virus infection.

Example 16 An in vivo activity study of dicycloplatin alone (orally or by injection) in patients with hepatitis B, hepatitis C, HIV infection and Ebola virus.

[111] An aqueous injection of dicycloplatin was purchased from Beijing Xingda Technology System Co., Ltd. and its concentration was 50 mg/5 ml. Dicycloplatin used in oral preparations is prepared with reference to US2016/0297842 and is obtained by dissolving dicycloplatin in deionized water.

[112] Experimental Methods: (1) Twelve HBV patients were given dicycloplatin separately for 1-3 months, the viral index and the liver function index were observed. Six of them were administered by intravenous drip, and 150 mg of an aqueous solution of dicycloplatin was dissolved in 250 ml of a 5% aqueous dextrose solution once every two days; 6 of them were administered orally, and 150 mg of an aqueous solution of dicycloplatin was dissolved in 250 ml of a 5% aqueous solution of dextrose. Once every two days. (2) Dicycloplatin was given to 9 hepatitis C patients separately for 1-3 months. Viral index and liver function were monitored by intravenous drip. 150 mg of an aqueous solution of dicycloplatin was dissolved in 250 ml of a 5% aqueous solution of glucose. It was taken once every two days; 6 of them were taken orally, and 150 mg of an aqueous solution of dicycloplatin was dissolved in 250 ml of a 5% aqueous glucose solution. It was taken once every two days. (3) Dicycloplatin was given to only 8 HIV-infected individuals (viral index >> 100,000 IU/mL) for 1-3 months (orally and by injection in four patients) to monitor viral index and liver function; dicycloplatin (reference to the above method, intravenous aqueous glucose solution) was given separately to 20 people infected with Ebola virus (Africa) for 1-3 weeks, and the viral index and liver function were observed.

[113] Results of the experiment: (1) Dicycloplatin was given to 12 patients with hepatitis B for 1-3 months. None of the patients showed a downward trend in viral index, while the viral index continued to rise and liver function did not improve; dicycloplatin was given separately to 9 patients with hepatitis C for 1-3 months. None of the patients showed a downward trend in viral index, while the viral index continued to rise and liver function did not improve; (3) 8 doses of dicycloplatin were given separately to 8 HIV-infected patients (viral index >> 100,000 IU/mL). After 1-3 months of use (oral and by injection in four patients), none of the patients showed a downward trend in the viral index, while the viral index continued to increase and liver function did not improve; Dicycloplatin was administered to 20 patients infected with Ebola virus for 1-3 weeks. None of the patients showed a downward trend in the viral index, and the viral index continued to rise, without any control or remission.

Example 17 Study of the in vivo activity of combination products containing dicycloplatin in patients with hepatitis B, hepatitis C and HIV infection.

[114] Experimental methods: (1) the combined product was applied to 15 patients with hepatitis B (without taking any other antiviral drugs), and after 1-3 months of treatment, the viral index and liver function index were observed; Of these, 6 were tested by taking capsules containing the combination products obtained in Experiment 4, which can be made according to conventional techniques known to those skilled in the art. As for the amount of dicycloplatin, it was taken once every two days at a dose of 150 mg of dicycloplatin; in 9 cases, the combined product obtained in experiment 10 was taken. As for the amount of dicycloplatin, it was taken once every two days in capsules at a dose of 150 mg of dicycloplatin; (2) the combination drug was used in 13 patients with hepatitis C (without any antiviral treatment). After 1-3 months of treatment, the viral index and liver function tests were monitored; 5 of them took the combined product obtained in experiment 7. With regard to the amount of dicycloplatin, it was taken three times a day in capsules at a dose of 50 mg of dicycloplatin; 8 of them used the combined product obtained in experiment 11. With regard to the amount of dicycloplatin, it was taken three times a day in capsules at a dose of 50 mg of dicycloplatin; (3) the combined product was used in 8 HIV-infected patients (without taking any other antiviral drugs), the viral index and liver function were monitored; 2 of them used the combined product obtained in experiment 4 three times a day. With regard to the amount of dicycloplatin, the dose in capsules is 50 m; 6 of them used the combined product obtained in experiment 11. With regard to the amount of dicycloplatin, they used three times a day in capsules at a dose of 50 mg of dicycloplatin.

[115] Results of the experiment. In this invention, it was disclosed that combination products containing dicycloplatin were used in 15 patients with hepatitis B (without taking any other antiviral drugs). After 1-3 months of treatment, the viral index decreased significantly, and liver functions were mostly normal; a combination preparation containing dicycloplatin was used in 13 patients with hepatitis C (without taking other antiviral drugs). After 1-3 months of treatment, the viral index decreased significantly. In 2 cases, the viral index fell below the detection limit, and the liver function index was mostly normal. The combination product containing dicycloplatin disclosed in this invention was used in 8 patients with HIV infection (without taking any other antiviral drugs). During treatment, the viral index decreased significantly, the CD ₄ index increased significantly, and liver function tests were mostly normal.

Example 18 Study of the in vivo activity of a combination product containing dicycloplatin in patients with hepatitis C and HIV infection.

[116] In the following cases (1) and (2), the combined capsule product obtained in experiment 6 was used, in which the capsules can be prepared according to conventional methods known to those skilled in the art, 150 mg of dicycloplatin capsules were taken per day ; in the following cases (3), (4) the combined capsule product obtained in experiment 10 was used, in which the capsules can be prepared according to conventional methods known to those skilled in the art. As for the dosage of dicycloplatin, 150 mg capsules were taken per day.

[117] Experimental Methods: (1) Hepatitis C patient Mr. He, aged 61, did not respond to anti-interferon and other antiviral drugs. The RNA virus index continued to rise. The RNA virus index was 472,000 IU/ml. He suffered from typical liver pain. One month after oral administration of the combined product containing dicycloplatin, the patient was subjected to the determination of the RNA virus index and liver function parameters; (2) Hepatitis C patient Mr. Wang, aged 51, without any treatment, has hepatitis C type III, whose RNA virus count was 932,000 IU/ml, and his liver showed typical pain and pressure. After taking the combined preparation containing dicycloplatin, the patient was monitored for pain and pressure in the liver for one month. Then two months later, the RNA virus index and the liver function index were checked; (3) A 43-year-old patient underwent a virological test and was confirmed to have HIV infection and clinical type C2. The test results showed that the number of CD ₄ lymphocytes was 312, CD ₄ cells was 319, and CD ₄ / CD ₈ = 0.58:1. Four months after oral administration of the combined product containing dicycloplatin, CD ₄ , CD ₈ cells and their ratios were detected; (4) The patient, 28 years old, underwent a virological test and was confirmed to have HIV, 190 CD ₄ cells, 815 CD ₈ cells, and CD ₄ / CD ₈ = 0.23:1. Six months after oral administration of the combined product containing dicycloplatin, CD ₄ , CD ₈ cells and their ratios were detected.

[118] Experimental results: (1) Hepatitis C patient, 61-year-old Mr. He, took the combination product containing dicycloplatin for one month, and the quantitative index of the RNA virus decreased from 472,000 IU/ml to 4,520 IU/ml. Liver function tests did not change significantly, and the patient's pain disappeared; (2) Hepatitis C patient Mr. Wang, aged 51, after taking a combination product containing dicycloplatin for one month, pain and pressure disappeared, and he continued to take it for two months. His RNA virus index dropped to below 1000 IU/mL and his liver function was mostly normal; (3) The 43-year-old female patient mentioned earlier was virologically HIV-positive and had clinical type C2. The test results showed that the number of CD ₄ lymphocytes was 312, CD ₄ cells was 319, and CD ₄ / CD ₈ = 0.58:1. Four months after oral administration of the combination product containing dicycloplatin, the test results showed the following: the number of CD ₄ cells was 568, CD ₈ was 606, and CD ₄ / CD ₈ = 0.92 1. (4) The patient, 28 years old, was HIV positive on virological examination and her CD4 cell count was 190, CD8 was 815, and CD ₄ /CD ₈ = 0.23:1. After six months of oral administration of the combination product containing dicycloplatin, the virological test was negative, the CD ₄ count was 563, CD ₈ was 575, and CD ₄ /CD ₈ = 0.97:1. The above results show that dicycloplatin has low direct antiviral activity. The results of using dicycloplatin or a combination product with dicycloplatin alone, obtained from patients infected with multiple viruses, show that dicycloplatin alone does not show clinical efficacy in antiviral treatment in patients. Dicycloplatin combination product is used to treat people infected with HCV, HBV and HIV, and the effect is remarkable.

Example 19 Stability study of combination products containing dicycloplatin.

[119] This invention discloses a combination obtained by mixing dicycloplatin with various acids. In order to achieve greater stability of dicycloplatin and try to improve its solubility, the present invention discloses the ratio of dicycloplatin and various acids to form a mixture, and methods for its preparation.

(1) Process for producing a mixture of dicycloplatin and 1,1-cyclobutanedicarboxylic acid

[120] Using a conventional preparation method, mixtures can be prepared by mixing dicycloplatin and a certain proportion of 1,1-cyclobutanedicarboxylic acid, otherwise by freeze-drying an aqueous mixture of dicycloplatin and a certain proportion of 1,1-cyclobutanedicarboxylic acid, both completely dissolved in water.

No change in the stability of dicycloplatin after mixing is determined by XRPD characterization and ¹ H-NMR.

[121] Combined mixtures can be obtained from carboplatin and a certain proportion of 1,1-cyclobutanedicarboxylic acid in water as a medium. The mixtures are stirred at 0-40° C. for 1 to 24 hours, then directly lyophilized and uniformly mixed to obtain the desired mixtures. Or mixtures can be obtained by direct lyophilization of a clear solution obtained by adding an appropriate amount of water to the above mixtures. The XRPD structure and ¹ H-NMR show that the mixture consisted of dicycloplatin and 1,1-cyclobutanedicarboxylic acid. No carboplatin residue was found. This method of production significantly improves the yield of dicycloplatin and allows complete consumption of raw carboplatin.

(2) Preparation of a mixture of dicycloplatin and citric acid, chlorogenic acid, citric acid, gallic acid, etc.

[122] Combined products can be obtained by conventional methods. It is fashionable to prepare by mixing dicycloplatinum and a certain proportion of one or more of 1,1-cyclobutanedicarboxylic acid, citric acid, chlorogenic acid, citric acid, gallic acid, ferulic acid, maleic acid, salicylic acid, etc. Or it can be prepared by carefully dissolving dicycloplatin and a certain proportion of one or more acids in water and subsequent lyophilization, the acids being 1,1-cyclobutane dicarboxylic acid, citric acid, chlorogenic acid, citric acid, gallic acid, ferulic acid and maleic acid, salicylic acid, etc. No change in the stability of dicycloplatin after mixing is determined by XRPD characterization and ¹ H-NMR.

(3) Preparation of dicycloplatin or its combination products with other components.

[123] Additional other components include coenzyme Q10, curcumin, glutathione (GSH), oryzanol, vitamin C, and anthocyanins.

[124] As shown in examples 5-10, the preparation of combination products using dicycloplatin as the main active ingredient can be carried out either by conventional methods or by a special preparation method for a particular situation.

[125] In many examples, dicycloplatin is obtained as a lyophilized powder and aqueous injection using water as a carrier. Stability experiments show that the composition retains good stability. In many instances, dicycloplatin has good compatibility and stability with appropriate pharmaceutical excipients and is thus formulated into suitable oral preparations.

Example 20 Stability study of dicycloplatin with 1,1-cyclobutanedicarboxylic acid and excipients

[126] The stability of the combined product containing dicycloplatin for eight consecutive weeks is shown in Tables 1-4 below.

[127] 1. The solid and solution product formed by dicycloplatin and 1,1-cyclobutanedicarboxylic acid (1:1.2 to 1:20 dicycloplatin in acid) are stable at 40°C. The above solid product was examined for eight consecutive weeks in/in the light, and the results showed that it was stable.

Table 1. Stability study of dicycloplatin with 1,1-cyclobutanedicarboxylic acid and excipients

Compositions
property Property and ratio of compositions Starting Qty
DCP(%) after 1 day
DCP(%) after 3 days
DCP(%) DCP(%) after 5 days In 2 weeks
DCP(%) DCP(%) after 3 weeks DCP(%) after 4 weeks DCP(%) after 5 weeks DCP(%) after 6 weeks DCP(%) after 7 weeks DCP(%) after 8 weeks Note Hard ( 40°C) Solid (1:1.2) 92 91 94 94 96 93 90 91 91 94 92 stable Solid (1:1.5) 86 86 85 82 86 84 84 83 87 89 88 stable Solid (1:3) 65 62 62 66 65 67 47 59 61 63 64 stable Solid (1:5) 40 38 42 45 44 43 39 40 39 40 42 stable Solid (1:10) 28 thirty 27 27 thirty 29 26 28 29 28 29 stable Solid (1:20) fifteen fifteen 16 16 16 16 fifteen fifteen fifteen 16 16 stable Water solution( 40°C) Aqueous solution (1:1.2) 92 90 91 93 93 90 88 88 90 89 90 stable Aqueous solution (1:1.5) 86 85 85 87 87 83 82 83 85 84 85 stable Aqueous solution (1:3) 65 63 61 62 62 58 58 59 62 61 63 stable Aqueous solution (1:5) 40 39 39 40 40 37 37 38 39 39 39 stable Aqueous solution (1:10) 28 thirty 29 thirty thirty 28 28 28 29 29 29 stable Aqueous solution (1:20) fifteen 16 16 16 16 fifteen fifteen fifteen fifteen 17 16 stable Solid
in the light Hard (1：1.2) 92 94 93 94 95 91 90 93 92 92 93 stable Hard (1:1.5) 86 86 89 86 84 83 85 86 88 85 88 stable Solid (1：3) 65 68 67 64 66 63 61 60 58 62 63 stable Solid (1：5) 40 44 45 40 43 42 40 41 41 42 43 stable Solid (1：10) 28 29 thirty 28 thirty 29 27 28 31 29 29 stable Hard (1：20) fifteen fifteen 16 16 16 fifteen fifteen fifteen 16 fifteen fifteen stable

[128] 2. The solid product formed by dicycloplatinum and acids such as vitamin C, gallic acid, salicylic acid, citric acid, ferulic acid, etc. was stable at 40°C.

Table 2. Stability of solid compositions formed by dicycloplatin and acids such as vitamin C, gallic acid, salicylic acid, citric acid, ferulic acid, etc.

Composition property Property and ratio of compositions Starting Qty
DCP(%) In 1
day
DCP(%) After 3
days
DCP(%) In 5
days DCP( %) DCP(%) after 2 weeks DCP(%) after 3 weeks DCP(%) after 4 weeks DCP(%) after 5 weeks DCP(%) after 6 weeks DCP(%) after 7 weeks DCP(%) after 8 weeks Note Hard ( 40°) DCP+VC(1:2) 98 97 95 100 101 100 99 96 99 99 100 stable DCP + gallic acid (1:2) 96 99 94 95 98 98 99 95 97 99 97 stable DCP + salicylic acid (1:2) 97 95 96 95 96 97 95 98 96 97 99 stable DCP + citric acid (1:2) 97 98 93 95 96 99 93 95 97 96 97 stable DCP + ferulic acid (1:2) 98 100 96 99 99 100 98 96 95 98 97 stable

[129] 3. The solution product formed by dicycloplatin and gallic acid, salicylic acid, citric acid, ferulic acid, etc., starts to decompose at 40°C after basically two weeks.

Table 3. Stability of the product in the form of an aqueous solution formed by dicycloplatin and gallic acid, salicylic acid, citric acid, ferulic acid,

composition property Compositions and ratio Starting Qty
DCP(%) after 1 day
DCP(%) DCP(%) after 3 days DCP(%) after 5 days DCP(%) after 2 weeks DCP(%) after 3 weeks DCP(%) after 4 weeks DCP(%) after 5 weeks DCP(%) after 6 weeks DCP(%) after 7 weeks DCP(%) after 8 weeks Note Water solution( 40°) DCP+VC(1:2) 98 90 84 84 70 61 58 58 57 57 56 Broke down after 2 days DCP + gallic acid (1:2) 96 94 96 97 92 81 76 72 70 63 58 Broke down after 2 weeks DCP + salicylic acid (1:2) 94 93 94 96 95 89 89 90 90 / / Broke down after 3 weeks DCP + citric acid (1:2) 97 94 90 92 91 84 86 85 87 88 87 Broke down after 2 days DCP + ferulic acid (1:2) 98 / / / / / / / / / / Broke down after 1 day

[130] 4. Mixing stability experiments of dicycloplatin with appropriate excipients prove that blends with one or more excipients are stable for 8 consecutive weeks at 40°C.

Table 4. Mixing Stability Experiments of Dicycloplatin with Appropriate Excipients

time/DCP( %) MD36792 + talcum powder MD36792+
Microscopic silicon powder MD36792+ Magnesium Stearate Note Dose 1g + 1g 1g + 0.5g 1g + 0.5g terms 40°C lighting 40°C lighting 40°C lighting stable Starting Qty 101 101 101 101 101 101 stable after 1 day 102 103 101 99 103 102 stable after 3 days 100 103 101 100 102 100 stable after 5 days 103 102 100 97 102 103 stable after 10 days 102 99 99 98 101 102 stable after 15 days 103 99 99 99 101 103 stable After 20 days 103 102 100 100 103 103 stable after 30 days 102 101 101 100 102 102 stable

Notes:

1. MD36792 refers to dicycloplatin;

2. The ratio refers to the molar ratio of dicycloplatinum (DCP) to 1,1-cyclobutanedicarboxylic acid;

3. Experimental conditions of high temperature T=40°;

4. Lighting experiment conditions were 2000 lux;

5. Solution concentration: 1 mg/ml;

6. The error is in the range of 2% up and down.

Example 21 Effect of Dicycloplatin on Pain and Inflammation

[131] The authors of this application have found that dicycloplatin, as the main active ingredient, has a significant effect in the treatment of pain in patients with cancer. Among cancer patients, it is especially suitable for patients with advanced liver cancer, kidney cancer, bone metastases, brain metastases and bone cancer. Patients can be treated with dicycloplatin injection or oral preparations as the sole treatment, as shown in Table 5. The inventors also found that a certain number of patients with advanced cancer did not show sensitivity to dicycloplatin and did not show tumor suppression effects. However, the apparent relieving effect of dicycloplatin on cancer pain is still evident and the pain even disappears. Some patients have stopped using dicycloplatin after a certain course of treatment with dicycloplatin and the pain has not recurred.

Table 5. The effect of dicycloplatin on pain as the main active substance

cases 5th ^day 17th ^day 31st ^day , then discontinuation ^{60s -90s}
(without using DCP) Effect on tumor suppression Lung cancer with bone metastasis 5 cases The pain is gone No pain No pain No pain No obvious changes after 4 weeks of treatment Bone cancer 3 cases The pain is gone No pain No pain No pain No obvious changes after 4 weeks of treatment Liver cancer
5 cases The pain is gone No pain No pain No pain No obvious changes after 4 weeks of treatment Lung cancer with brain metastasis
2 cases The pain is gone No pain No pain No pain 1 case: no obvious change after 4 weeks of treatment
1 case: somewhat effective kidney cancer
1 case Pain eased The pain is gone No pain No pain No obvious changes after 4 weeks of treatment

[132] The above cancer patients were dosed once every two days, via intravenous drip of 150 mg/5% aqueous glucose solution. The dicycloplatin injection used was purchased from Beijing Xingda Technology System Co., Ltd. The concentration was 50 mg/5 ml.

[133] In the clinical treatment of tumor patients, dicycloplatin has been found to have a good effect on tumor-induced edema and tissue inflammation. In many cases of patients with brain metastases from lung cancer who took dicycloplatin orally or by injection, it was found that after one to two weeks of treatment with dicycloplatin, tissue edema caused by cancer metastasis to the brain was largely eliminated. This shows that dicycloplatin has a good anti-inflammatory effect in inflammation. In many cases of liver cancer patients with edema of the liver tissue, after infusion of dicycloplatin or oral administration for a certain period of time, the edema of the tissue disappeared or decreased significantly.

Example 22 Antivirus effects of other components AA011 and VK021 separately

[134] AA011 and VK021 information is presented in Table 6.

sample Molecular mass Weight
(mg) +DMSO(µl) Initial concentration (mc)M Initial concentration (µg/ml) dilution gradient AA011 309 16.4 531 1000 324 4 x multiple VK021 309 thirty 971 1000 324 4 x multiple

[135] As shown in FIG. 5, the antiviral effects of AA011, VK021 and positive drugs are shown in the figure below. The positive 2'CMC drug has a good antiviral effect, and the antiviral effect is better with increasing drug dose. Supplement AA011 has a definite antiviral effect at 1000 µM and 250 µM, and at 1000 µM there is no apparent cytotoxicity. The VK021 supplement also had some antiviral effect at 1000 µM and 250 µM, but had weak cytotoxicity at 1000 µM.

Example 23 In Vitro Efficacy of MD36792 and Combined Formulations Containing MD36792, AA011 and VK021 against HCV

[136] Sample MD36792, AA011 and VK021 and test them as shown in Table 7.

sample Molecular mass Initial concentration dilution gradient CC50 EC50 SI MD36792 500 200mcg/ml 4X 69.4 15.62 4.44 MD36792 500 200mcg/ml 4X 24.82 4.43 5.60 AA011 309 4000mcg/ml VK021 309 4000mcg/ml

[137] The antiviral effect of MD36792, AA011 and VK021 was shown in Fig.6.

[138] The antiviral effect of the combination of MD36792 and AA011 was shown as A in Fig. 6, while MD36792 and VK021 were shown as V in Fig. 6. The EC50 for MD36792 alone is 15.62µg/mL, the toxic CC50 for cells is 69.4, and the antiviral effect SI selection index is 4.44. The selection index is relatively low, which can be considered a certain antiviral effect, but the toxicity to cells is relatively high. The antiviral effects of MD36792 in combination with AA011 and VK021 are EC50 equal to 4.43 μg/ml, CC50 equal to 24.82 and SI equal to 5.6. The mixed use of three samples is more toxic to the cells, and the selection index is not very high. The antiviral effect of the combined use of MD36792 alone at the cellular level did not improve significantly.

Example 24 Study of analgesic effect of MD36792

[139] (I) Experimental materials, animals and equipment

Animals: 50 ICR mice, 18-22 g (Changsha Sleck Jingda Experimental Animal Company)

Reagent: rotundin (manufactured by Guangzhou Baiyunshan Pharmaceutical).

instrument: glass jar

Drug: MD36792

[140] (II) Experimental methods and experimental results

[141] Experimental method: animals were fed adaptively for 5 days and the body weight of mice was measured. Using stratified random division into groups, 50 mice were randomly divided into 5 groups, distilled water was used as a blank control group, ie. model group, Rotundin used for the positive control group referred to 100mg/kg, 1mg/ml; the aqueous solution of MD36792 used for the low dose group referred to 10mg/kg, 2mg/ml; the aqueous solution of MD36792 used for the middle dose group referred to 20mg/kg and 4mg/ml; the aqueous solution of MD36792 used for the high dose group referred to 40 mg/kg. Mice assigned to the MD36792 group and the positive control group were given the appropriate drugs. 1 hour after administration, mice in each group were intraperitoneally injected with 0.8% acetic acid (0.2 ml/individual). The amount of twisting of the mice was observed and recorded for 15 minutes.

Table 8

Group Sample analysis Number of mice Dose(mg/kg) weight Number of twists P Mean SEM Mean SEM A Model group ten 0 23.9 0.5 19.0 2.1 B Rotundin group ten 100 23.9 0.3 2.8 1.2 0.000** C Low dose MD36792 ten ten 23.5 0.5 15.2 3.6 D Average dose MD36792 ten twenty 24.0 0.4 14.8 2.2 E High dose MD36792 ten 40 23.6 0.4 8.7 3.1 0.009**

*Compared to the model group (i.e. the blank control group), there is a statistically significant difference, P < 0.05 **

*Compared to the model group (i.e., the blank control group), there is a statistically significant difference, P < 0.01

[142] Within 15 minutes after intraperitoneal injection of 0.8% acetic acid, the average number of twists in the model group reached 19 times. Compared with the model group, the high-dose MD36792 group (40mg/kg) and the positive control group (rotundin, 100mg/kg) statistically significantly suppressed the number of twists in mice (P = 0.009 or P = 0.000, respectively) ). MD36792 is dose independent in terms of analgesic effects as shown in FIG. 7.

[143] Therefore, the classical model induced by acetic acid was used to evaluate the analgesic effect of drugs. The above data derived from the model showed that both MD36792 and the positive control drug Rotundin had significant analgesic effects.

Claims (13)

1. A method for treating and/or preventing pain, comprising administering an effective dose of dicycloplatin to a patient in need thereof. 2. The method of claim 1 wherein the patient is a cancer patient. 3. The method of claim 1, wherein the patient is a liver cancer patient, a kidney cancer patient, a bone metastasis patient, a brain metastasis patient, or a bone cancer patient. 4. The method of claim. 1, wherein the patient is a patient with advanced cancer. 5. The method of claim 1, wherein the patient is a patient who has already developed resistance to a platinum-based anticancer drug. 6. The method of claim 1 wherein the patient is a patient with advanced liver cancer. 7. The method according to any one of paragraphs. 1-6, wherein the effective dose of dicycloplatin for treating and/or preventing pain is lower than the effective dose of dicycloplatin for treating cancer in said patient. 8. The method according to any one of paragraphs. 1-7, in which dicycloplatin is the only active ingredient for the treatment and/or prevention of pain. 9. The method according to any one of paragraphs. 1-7, further comprising administering to the patient one or more additional active ingredients having an analgesic effect. 10. The method according to any one of paragraphs. 1-9, further comprising administering to the patient one or more anti-cancer active ingredients, wherein the one or more anti-cancer active ingredients are selected from non-platinum-based anti-cancer drugs. 11. The method of claim 1 wherein the patient is not a cancer patient. 12. The method of claim 11 wherein dicycloplatin is the sole active ingredient for the treatment and/or prevention of pain. 13. The method of claim 11 further comprising administering to the patient one or more additional active ingredients having an analgesic effect.

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