KR20130027576A - Compositions and methods of reducing tissue levels of drugs when given as orotate derivatives - Google Patents

Abstract

The present invention is in the field of chemical readjustment of drugs known to cause tissue toxicity as a side effect by introducing orotate derivatives of drugs. In particular, the present invention relates to orotetate derivatives of anthracycline, doxorubicin and daunorubicin that have been found to reduce levels of the drug in non-cancerous tissues. The orotate derivatives are equally effective at inhibiting subcutaneous CAKI-1 kidney tumors in animals and reducing the cardiac tissue of doxorubicin in comparison to doxorubicin hydrochloride and are toxic caused by free radical production by anthracyclines. Implies a decrease in

Description

COMPOSITIONS AND METHODS OF REDUCING TISSUE LEVELS OF DRUGS WHEN GIVEN AS OROTATE DERIVATIVES}

The present invention converts orthoacid derivative pharmaceuticals, especially cancer drugs, into their orotate derivatives, thereby making it possible for non-cancerous tissues (tissues not cancerous) that are susceptible to cancer drug toxicity. To reduce the tissue level of the drug.

The present invention relates to the field of chemical restructuring by producing their orotate derivatives of drugs known to cause toxic or adverse drug reactions in non-cancerous tissues as side effects. In particular, the present invention relates to derivatives of anthracyclines, doxorubicin and daunorubicin used as anticancer drugs.

Antibiotic doxorubicin (DOX) and derivatives thereof, like other cationic anthracyclines, are currently of great clinical interest in the treatment of cancer, including leukemias and solid cancers.

There is great hope for the use of liposomes as delivery systems for bioactive agents. In many cases it has been established that the use of liposomes for the administration of anti-neoplastics improves traditional methods of administration. Gavizon and his colleagues (Gabizon et al), Cancer Res. (1982), 42; 4234 to 4739; And Cancer Res (1984) 44 by Van Hossel et al. (Van Hossel et al); See 3698-3705. Other patents describe the inclusion of anti-free radical agents into liposomes with improved activity as inhibitors of lipid peroxidation. See US Patent No. 5,605,703, issued February 25, 1997. Liposomal encapsulation can substantially affect the functional properties of a drug compared to an unencapsulated drug. In addition, other liposome drug products may differ from one another in the chemical composition and physical form of the liposome. These differences can substantially affect the functional properties of liposome drug products. Doxorubicin HCL is (85, 105) -10-[(3-amino-2,3,6-tridioxy-alpha-L-lyxo-hexopyranosyl) oxyl-8-glycol-7 , 8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12-naphthacedione hydrochloride ((85, 105) -10-[(3-amino-2 , 3,6-trideoxy-α-L-lyxo-hexopyranosyl) oxyl-8-glycolyl-7,8,9,10-tetrahydro-6,8, ll-trihydroxy-l-methoxy-5, 12-naphthacenedione hydrochloride Name for. The molecular formula of the drug is C ₂₇ H ₂₉ NO ₁₁ HCl; Its molecular weight is 579.99.

Doxyl? (Doxorubicin hydrochloride injection) is doxorubicin hydrochloride encapsulated in Stealth® liposomes for intravenous administration. Private room above said stealth? Liposomes are composed of surface-bound methoxypolyethylene glycol, a process often referred to as pegylation, so that liposomes protect liposomes from detection by the mononuclear phagocyte system and increase blood circulation time. do.

Stealth? Liposomes have a half-life of approximately 55 hours in the human body. They are stable in the blood and direct measurement of liposome doxorubicin has shown that at least 90% of the drug remains liposome-encapsulated during circulation.

It is hypothesized that experiments at high cumulative doses are limited and therefore doxyl® may have a similar myocardial toxicity than the usual composition of doxorubicin hydrochloride. When the total dose of doxorubicin hydrochloride reaches 550 mg / m 2, one may face irreversible myocardial toxicity, often causing congestive heart failure that does not respond to cardiac support therapy. Swelling, headache, chills, back pain, stuffiness in the chest or neck, and / or hypotension occurred in more than 10% of patients treated with a single room. In most patients these responses resolve over several hours to a day once the infusion is complete. In some patients the response is resolved by slowing the infusion rate. Serious and sometimes life-threatening or fatal allergic reactions have been reported.

Doxorubicin has been successfully administered by extensive schedules, and the anti-tumor activity of doxorubicin is proportional to the area under the concentration * time curve (AUC), not the maximum drug levels. The AUC for concentrated doxorubicin hydrochloride is 9.9 mg / ml-h. The AUC (mg / ml-h) for Toxyl® is 590.

Cardiac toxicity ( cardiac toxicity )

The cardiotoxicity exhibited by doxorubicin and other anthracyclines is unique in terms of its pathology and mechanism. Key limitations in the clinical use of anthracycline in adults are bone marrow suppression, mucositis and drug resistance to the site of the tumor. However, in individual patients, cardiotoxicity can develop while most often the patient's tumor still responds to the drug for the use of doxorubicin to treat breast cancer. This is not only for use with anthracyclines or in combination with other chemotherapeutic agents, but also for the treatment of monoclonal antibodies trastuzumab and the advanced breast cancer itself. Is a problem with the use of antibodies directed directly against HER2 / non oncoprotein that is active in humans. The observed potential of anthracycline-induced heart damage by trastuzumab is high in tumors. It was eliminated by its use with doxorubicin in the distribution of patients exhibiting levels of HER2 / new expression. Young children are likely to be more sensitive to the cardiotoxicity of the drug, which has become a clear problem in the use of doxorubicin in pediatric oncology. See Management of Drug Toxicity, Ch 31-42, in The Chemotherapy Source Book, 3 ^rd ed, Michael C. Perry, Lippincott Williams & Wilkins, 2001.

Thus, there is a great need for the development of analogues that give better response rates, broader response bands and / or reduced cardiotoxicity. Halogenated anthracyclines are mechanically different from doxorubicin, and daunorubicin has been produced. In particular, derivatives with fluorine groups attached to their sugar moieties have a strong ability to kill tumor cells. Most of the origin and prior art of doxorubicin are found in published patents and published literature. United States Patent No. 5,304,687, issued April 19, 1994, Issue 5,605,703 issued February 25, 1997, Issue 6,210,930, issued April 3, 2001, issued September 4, 2001. See 6,284,737 and 6,653,455, issued November 25, 2003.

However, none of the prior art approaches the problem of inhibiting and / or reducing the level of drug in non-cancerous tissues as a strategy for reducing toxicity and drug side effects. It is distinguished. More effective and less toxic agents are being pursued more broadly and are a fundamental object of the present invention. Related points of the foregoing references are specifically cited herein by reference.

The present invention provides for the misuse of drugs that exhibit increased clearance of drugs from non-cancer tissues that are targets for drug toxicity when compared to non-derivatized forms of the drug. It seeks to overcome the disadvantages inherent in the prior art by providing compositions of rotate derivatives.

The present invention is in the field of chemical readjustment of drugs known to cause tissue toxicity as a side effect by introducing orotate derivatives of drugs. In particular, the present invention relates to orotate derivatives of anthracyclines, doxorubicin and daunorubicin, which are used as anticancer drugs.

In view of the foregoing techniques, the inventors have described a chemical organic moiety that reduces the cardiac tissue levels of the anthracycline of Formula A in FIG. 1, as illustrated by doxorubicin orotate and daunorubicin orotate. orotate derivatives of anthracyclines, including moiety).

It is a primary object of the present invention to obtain a composition of doxorubicin or daunorubicin, their orotate derivatives and analog compounds in order to reduce the toxicity of the drug, which is related to the tissue levels of the drug accumulating in the heart.

The present invention also provides a process for the preparation of doxorubicin orotate and related derivatives, especially starting with doxorubicin hydrochloride, orotic acid and potassium hydroxide. This process includes a) reacting potassium hydroxide with orotic acid, extracting potassium orotate, and reacting the extracted potassium orotate with doxorubicin hydrochloride to form doxorubicin orotate. do.

Another object of the invention is to treat human neoplasms and in particular primary or metastatic tumors, proliferative hematopoietic disorders and leukemia with doxorubicin orotate and given doxorubicin A method of reducing the drug's toxic secondary effects by reducing the level of the drug in non-cancer tissues, which are targets sensitive to drug toxicity, by 10 to 100% compared to hydrochloride. It is.

The present invention provides a method of reducing the level of a drug in non-cancer tissues, the method comprising the steps of: a) converting the drug into an orotate derivative composition, administering the orotate derivative to its required subject And measuring the level of the drugs in non-cancer tissues, wherein the drugs are acetanilide, actinomycin D, adriamycin, ami Noacridines, aminoimidazoles, aminoquinolines, anilides, anilides, anthracycline antibiotics, antiestrogens, benzazepines, benzhydryl compounds benzhydryl compound, bezodiazpine, benzofuran, cannabinoid, cephalosporine, cisplatin, kohl Colchicines, cyclic peptides, cyclophosphamides, daunorubicin, dibenzazepine, digitalalis glycosides, dihydropyridine, Doxorubicin, epiphodophyllotoxin, epirubicin, ergoline, ergot alkaloid, etoposide, 5-fluorouracil ), Idarubicin, ifosamide, imidazole, interleukin-2, alpha-interferon alpha, isoquinoline, macrolide , Melphalan, methotrexate, mitomycin-C, mitoxantrone, naphthalene, nitrogen mustard, opioid, oxazine (oxazine), oxazole, paclitaxel (p aclitaxel, phenothiazine, phenylalkamine, phenylpiperidine, piperazine, piperidine, polycyclic aromatic hydrocarbons, pyridine ), Pyrimidine, pyrrolidine, pyrrolidinone, quinazoline, quinoline, quinine, lauwolfa alkaloid, retinoid , Salicylate, steroid, stilbene, sulfone, sulfonylurea, tamoxifen, taxol, taxotere, THP-Adriamycin (THP-adriamycin), trastuzumab, triazole, tropane, tropane, vinblastine, vincristine or vinca alkaloid Will be.

In some embodiments of the present invention, the anthracycline orotate derivatives have the general formula of

Wherein R ₁ represents a hydrogen group (-H), a hydroxyl group (-OH), a methoxy group (-OCH ₃ ), an aryl group having 6 to 20 carbon atoms, or -O-CO- (CH ₂ ) _n CH Fatty acyl group having the general formula of ₃ , wherein n is an integer from 1 to 20, or fat having the general formula of -O-CO- (CH ₂ ) ₁ (CH = CH) _m (CH ₂ ) _n CH ₃ An acyl group, where l is an integer from 1 to 3, m is an integer from 1 to 6 and n is an integer from 1 to 9;

Each of R ₂ and R ₃ is independently a hydrogen group (—H), a hydroxyl group (—OH), a methoxy group (—OCH ₃ ) or a double bonded oxygen moiety;

R ₄ is a hydrogen group (-H), a hydroxyl group (-OH), a methoxy group (-OCH ₃ ) or a halide; And

Y ₁ and Y ₂ are each independently a hydrogen group (—H), a hydroxyl group (—OH), a methoxy group (—OCH ₃ ) or a double bonded oxygen, sulfur or nitrogen group.

The present invention converts orotic acid derivative drugs, especially cancer drugs, to their orotate derivatives to reduce the tissue level of the drug in non-cancerous tissues (tissues not cancerous) that are susceptible to cancer drug toxicity. It can be used to provide a formulation and the like that can be.

1 is a diagram showing the structure of daunorubicin and doxorubicin and their orotate derivatives.
Fig. 2 shows the synthesis of doxorubicin orotate.
3 is a mass spectrometry graph showing doxorubicin orotate.
4 is a nuclear magnetic resonance spectroscopy graph showing doxorubicin orotate.
5 is a graph showing the response of SC CAKI-1 kidney tumors treated with doxorubicin hydrochloride.
Figure 6 is a graph showing the response of SC CAKI-1 kidney tumors treated with doxorubicin orotate.
7 is a graph showing the response of SC CAKI-1 kidney tumors treated with doxorubicin hydrochloride and doxorubicin orotate.

Drug therapies used for the treatment of patients suffering from cancer can damage many organs and organ systems. The most frequent of these are tissues with rapid cell turnover, such as the hematopoetic system, the gastrointestinal tract, and the genitourinary tract. Hearts that are made of tissues that do not have rapid cell turnover and therefore cannot recover quickly are often affected by chemotherapy. Since the cardiac effects of these therapies can be disabled or life threatening, major changes in treatment may be inevitable to prevent cardiac toxicity. In some cases, the effects of chemotherapy drugs on the heart are self-limited and easily reversible with the recovery of the aggressive agent. In other cases, however, the damage can be destructive, gradual, irreversible and extremely fatal. Some forms of cardiotoxicity are rarely foreseeable and can sometimes affect a patient without warning even during the first exposure. In other settings, the toxicity is well defined and easily detectable. However, there are still significant changes in the exposure required to achieve similar levels of tissue damage.

Some drugs are toxic in themselves, but their toxicity may be increased when they are used in combination with other agents, the combination may be more toxic than the sum of the toxicities of the individual components. As it is necessary to achieve the maximum antitumor potential of the drug while maintaining terminal organ toxicity at an acceptable level, the assessment in patients treated with drugs of toxicity should be individualized.

The most cardiotoxic drugs are according to their cardiac effects: 1) drugs associated with myocardial depression (eg, anthracycline-doxorubicin, THP-Adriamycin, idarubicin, epirubicin, daunorubicin; Other anthraquinones-toxicity intensifiers such as mitoxanthrone, cyclophosphamide, mitomycin-seed, etoposide, melphalan, vincristine or bleomycin and other agents-alpha-interferon and trastuz Mab); 2) anticancer agents associated with anemia (eg 5-fluorouracil, vinblastine, vincristine, cisplatin or interleukin); 3) anticancer agents associated with hypotension (eg interleukin-2) and 4) various agents with cardiac toxicity (eg paclitaxel, actinomycin di, mitomycin C or ifosamide). have.

The most widely used doxorubicin in the anthracycline group has been overstudyed and functions as a model for drugs associated with cardiotoxicity and myopathies. Its effectiveness is in part limited by cumulative dose-related cardiotoxicity, which usually occurs late during the course of treatment, or may exist over several rows or years after completion of treatment. This strongly suggests that there may be drug residues left in the tissue after the treatment is completed. Thus, there is an absolute need to reduce and / or prevent the accumulation of drugs in the short and long term as well as immediately after the end of treatment in non-cancerous or non-disease tissues.

In particular, the presentation of toxicity is slowly gradual, potentially irreversible and life-threatening. The pathogenesis of doxorubicin-associated cardiomyopathy is not fully understood and, as well as the exact mechanism of cardiac toxin in patients treated with doxorubicin and related drugs, causes a series of cardiac dysfunctions. The cases are still under investigation. However, there is no question that the accumulation of doxorubicin and related drugs in heart tissue causes progressive cardiac dysfunction and damage, and therefore the object of the present invention is to accumulate drugs and their metabolites in target organs such as the heart. To reduce and / or prevent them. Doxorubicin has been shown to cause direct damage to the membranes as well as indirect damage via oxygen-free radicals that inhibit calcium transport and eventually inhibit cardiac function. Therefore, it is absolutely necessary to reduce or completely prevent the accumulation of doxorubicin in cardiac tissue to reduce potential short and long term side effects of the drug.

Doxorubicin cardiomyopathy can appear months and even years after successful completion of the course of chemotherapy at or near the maximum tolerated dose. Because a large number of patients treated with doxorubicin are treated and survive for long periods after their chemotherapy, some patients with obvious subclinical heart damage are rarely tolerated and months or years after initial cardiac insult You may experience separate stresses that cause symptoms of dysfunction. It is an object of the present invention to provide an initial treatment caused by the drug by reducing the level of the drug in the heart by preventing and / or rapidly safeguarding the accumulation of the drug from heart tissue when chemotherapy is applied. To reduce the size and incidence of heart damage.

In animal models, it has been observed that encapsulation of doxorubicin in liposomes significantly reduces the secondary effects of chronic and acute toxicity. Cancer Res (1980) 40 by Rahman et al (Rahman et al); 1532-1537, J. Natt Cancer Inst (1986) 77 of Gabzon and his colleagues (Gabizon et al); See 459-467. Other indicators of toxicity, such as alopecia, weight loss, nausea, vomiting and dermal necrosis by extravasate, can be markedly reduced by administration of doxorubicin in liposomes. . Cancer Treat Rep (1983) 67 from Forssen et al .; See 481-484. Importantly, it has been established in several cancer models that this marked reduction in toxicity does not reduce anticancer efficiency.

Cardiomyopathy observed in treatment with doxorubicin is similar to the lesions seen in cardiac muscle in experimental animals under conditions of alpha-tocopherol deficiency. These results suggest that the damage produced by the drug is caused by an increase in free radical reactions that are believed to involve the membrane of lipids. Thus, the inclusion of doxorubicin, referred to hereinafter as DOX, into the lipid bilayers of liposomes, facilitates free radical reactions. Thus, the levels of free radicals in DOX exposed non-cancerous tissues were measured by biochemical methods, demonstrating that orotate derivatives of DOX lower their levels.

The object of the present invention is to be caused by the drug by reducing the level of the drug in the heart by preventing and / or rapidly safening the accumulation of the drug from the heart tissue when chemotherapy is administered. To reduce the size and incidence of early heart damage.

Orotate  Properties of Drugs Used as Derivatives

As used herein, the word "drug" is defined as a chemical designed to be used in the treatment or prevention of a disease. Drugs include recognized synthetic and naturally occurring bioaffecting substances as listed in “The Physician desk Reference,” 56th ed, pages 101-133 (or updated editions). The term “drug” also includes compounds having marked properties that are not found or obtainable. The present invention can be used with drugs consisting of charged, uncharged, hydrophilic, zwitter-ionic or hydrophobic species and likewise combinations of these physical properties. Hydrophobic drugs are defined as drugs whose non-ionized form can be better dissolved in lipids or fats than in water. Preferred classes of hydrophobic drugs are drugs that are more soluble in octanol than water.

Compounds or drugs selected from various classes of compounds may be converted into orotate derivatives and administered orally as orotate derivatives. Such compounds or drugs include, for example, the following classes: acetanilide, actinomycin di, adriamycin, aminoacridine, aminoimidazole, aminoquinoline, anilide, anthracycline antibiotics, antiestrogens, benzase Pin, benzhydryl compound, benzodiazepine, benzofuran, cannabinoid, cephalosporin, cisplatin, colchicine, cyclic peptide, cyclophosphamide, daunorubicin, dibenzazepine, digitalis glycoside, dihydropyridine, doxorubicin , Epipodophyllotoxin, epirubicin, ergoline, ergot alkaloid, etoposide, 5-fluorouracil, idarubicin, ifosamide, imidazole, interleukin-2, alpha-interferon, iso Quinoline, Macrolide, Melparan, Methotrexate, Mitomycin-C, Mitoxantrone, Naphthalene, Nitrogen Mustard, Opioid, Oxazine, Jade Sol, paclitaxel, phenothiazine, phenylalkamine, phenylpiperidine, piperazine, piperidine, polyaromatic hydrocarbons, pyridine, pyrimidine, pyrrolidine, pyrrolidinone, quinazoline, quinoline, quinine, lauulpa Alkaloids (rauwolfa alkaloid), retinoids, salicylates, steroids, stilbenes, sulfones, sulfonylureas, tamoxifen, taxol, taxotere, THP-Adriamycin, trastuzumab, triazole, tropanes, vinblastine, vincacristine Or vinca alkaloids, including but not limited to these.

"Side effects" or "toxicity" or "drug side effects" of chemotherapeutic agents are observed in the acute state of chemotherapy administration and in patients treated with cancer with subclinical tissue damage. There is a high awareness of drug-related tissue side effects that can be quite severe, disabled and irreversible. The clinician should be aware of potential tissue / organ complications of chemotherapeutic agents and where baseline tissue examination should be appropriately performed prior to initiating the therapy described above.

The "clearance" of a drug occurs by the perfusion of blood to the organ of extraction. "Extract" means the ratio of drug to organ that has been irreversibly removed (secreted) or changed (metabolized) to another chemical form. Clearance (CL) is therefore calculated as the product of the ratio of blood flow through the trachea and the drug extracted by the trachea.

The clearance of the drug usually arises from the liver and kidney, and it is assumed that only free and unbinded drugs are obtainable for clearance. For liver clearance, hepatocellular membranes obtainable as lipophilic drugs by sinusoidal carrier systems for specially ionized molecules (anionic and cationic) of molecular weights of 400 or more Passive diffusion through the lipid core of hepatocyte membranes is enhanced. Similarly, other transporters on the canalicular face transport drugs or metabolites into the bile. This system has two separate processes: hepatic uptake and billiary excretion. For small sized lipophilic drugs that easily cross the membranes, intrahepatic migration is not the major factor, but for compounds containing higher molecular weight compounds (greater than 500) and significant H-bonding In contrast, even if metabolism subsequently occurs, intrahepatic migration can be a key clearance process.

The present invention provides a method of increasing the clearance of orotate derivatives of the drug from non-cancer tissues by at least 25% relative to the dosage of the drug as measured by pharmacological studies. The invention also provides a method of increasing the clearance of orotate derivatives of the drug from non-cancer tissues by at least 50% relative to the dose of the drug as measured by pharmacokinetic studies. The present invention also provides a method of increasing the clearance of orotate derivatives of the drug from non-cancer tissues by at least 100% relative to the dose of the drug as measured by pharmacological studies.

The present invention provides a composition that increases the clearance of orotate derivatives of the drug from non-cancer tissues by at least 50% relative to the dosage of the drug as measured by pharmacological studies. The invention also provides compositions that increase the clearance of the orotate salt of the drug from non-cancerous tissues by at least 100% relative to the dose of the drug as measured by pharmacokinetic studies. The present invention provides a composition that increases the clearance of orotate derivatives of the drug from non-cancer tissues by at least 100% relative to the dose of the drug as measured by pharmacokinetic studies.

Absorption or efflux occurs by any of three methods: passive diffusion, active transport, or facilitated active transport. Passive diffusion is a simple passage of molecules across the mucosal barrier until the concentration of molecules reaches an osmotic balance on both sides of the membrane. In active transport, molecules are actively pumped across the mucosa. In facilitating active transport, carriers, which are generally proteins, are required to transport molecules across the membrane for absorption.

How to reduce the side effects of a formulation by converting it to an orotate derivative

In co-pending application No. 11/063943, filed February 22, 2005, the inventors have found an oral bioavailability of a medicament by converting the medicament, which is hardly absorbed from the gastrointestinal tract, into orotate salts. It describes how to increase bioavailability. In the present invention, the inventors have reduced levels in the organs of drugs given with orotate derivatives as compared to pharmaceutical forms of the drug, and thus at the time of drug administration and for a long time after the primary cancer or disease has been treated. To reduce the potential for toxicity. Thus, particularly useful compositions of orotate derivatives of a medicament can provide rapid onset and sustained action using lower dosages and reduce drug interactions and side effects. All referenced references are hereby fully incorporated by reference.

Free pyrimidine (orotic acid) is important in the synthesis of urpylate (UPP), the main pyrimidine nucleotide. Pyrimidine plays a central role in cellular regulation and metabolism. These are substrates for DNA / RNA biosynthesis, regulators of the biosynthesis of some amino acids and cofactors in the biosynthesis of phospholipids, glycopeptides, sugars and polysaccharides. The traditional de novo pyrimidine biosynthetic pathway is terminated by the synthesis of UMPs. Biochemistry, ed Lubert Stryer, ed, W.H. See Freeman & Co NY, 4th ed, 739-762 (1995). It has been reported that 5-Fluorouracil is toxic as measured by acid soluble fraction, inclusion in RNA and DNA in normal tissues in mice's liver. Orotic acid administration reduces the inclusions in the liver and intestinal RNA, thus suggesting a decrease in 5-fluorouracil induced toxicity in the liver. See In vivo 1: 309-312 (1987) by El Hag IA et al. The present invention provides drug orotate derivatives that proceed to dissolution to release the drug as charged molecules and free orotic acid, which in turn reduces drug-induced liver, heart or other tissue toxicity.

The present invention provides methods and compositions for increasing the effectiveness of a drug by converting the drug into an orotate derivative composition and administering the orotate derivative composition to a subject in need thereof, wherein the drug is acetanilide, actinomycin. Di, adriamycin, aminoacridine, aminoimidazole, aminoquinoline, anilide, anthracycline antibiotic, antiestrogen, benzazepine, benzhydryl compound, benzodiazepine, benzofuran, cannabinoid, cephalosporin Cisplatin, colchicine, cyclic peptide, cyclophosphamide, daunorubicin, dibenzazepine, digitalis glycoside, dihydropyridine, doxorubicin, epipodophyllotoxin, epirubicin, ergoline, ergoline alkaloid , Etoposide, 5-fluorouracil, idarubicin, ifosamide, imidazole, inter Kin-2, alpha-interferon, isoquinoline, macrolide, melfaran, methotrexate, mitomycin-c, mitoxanthrone, naphthalene, nitrogen mustard, opioid, oxazine, oxazole, paclitaxel, phenothiazine, phenyl Alkamine, phenylpiperidine, piperazine, piperidine, polyaromatic hydrocarbons, pyridine, pyrimidine, pyrrolidine, pyrrolidinone, quinazoline, quinoline, quinine, lauwolfa alkaloid, retinoid, salicylic acid Selected from the group consisting of salts, steroids, stilbenes, sulfones, sulfonylureas, tamoxifen, taxol, taxotere, THP-adriamycin, trastuzumab, triazole, tropane, vinblastine, vincacristine or vinca alkaloids do.

The present invention is directed to a method for treating a drug as determined by reducing organ / tissue levels of the drug in cases where the drug is known to cause toxicity or has the potential to induce toxicity in long-term tissue accumulation of the drug. Methods and compositions for increasing the effectiveness of rotate derivatives are provided.

6. Examples

Example  One.

Doxorubicin Orotate  Chemical synthesis

2 shows the synthesis of doxorubicin orotate. Orotic acid (1.0 g) was dissolved in 200 mL of water at 65 ° C. Potassium hydroxide (KOH) (1 equiv) was added to the orotic acid and the mixture was stirred at 85 ° C. until a clear solution appeared. The solution was cooled and the precipitate collected by filtration to give a colorless solid (0.95 g), which was dried in vacuo for 24 h. Mass spectroscopy indicates that the structure is potassium orotate.

Potassium orotate synthesized in the previous step (0.4 g) was dissolved in 150 ml of water at 65 ℃. The solution was degassed in vacuo and protected with argon. Doxorubicin hydrochloride (1.0 g, 1 equiv) was added and the resulting red solution was treated with excess amberlite IR 120 H resin for 2 hours at the same temperature. After filtration the solution was frozen in dry ice and lyophilized under reduced pressure to yield 1.35 g of doxorubicin orotate (J-I220-13-II) as a red solid. Mass spectrometry (FIG. 3) and nuclear magnetic resonance (FIG. 4) indicate that the structure is doxorubicin orotate.

Example  2.

SC for treatment with doxorubicin hydrochloride and doxorubicin orotate Response of CAKI- 1 Renal Tumor

The purpose of this experiment was to subdoxically subcutaneously (SC; subcutaneously) male SCN-1 male kidneys into doxorubicin hydrochloride (DOX) for human kidney tumor xenografts. ) And its orotate derivative (DOX orotate) to evaluate the anticancer effect. Hearts of animals treated with equivalent doses of DOX and DOX orotate were analyzed and the concentration of DOX in the hearts of these animals was measured.

Drug Composition—Composed in fresh saline each day treated with a 0.8 mg / ml solution of DOX (doxorubicin hydrochloride USP28, Yick-Vic Chemical & Pharmaceuticals (HK) Ltd, batch number MO5O7O5, Kowloon, Hong Kong). The 0.8 mg / ml solution was diluted to 0.53 and 0.35 mg / ml with brine.

As described in Example 1, DOX orotate (lot number J1220-13-II) was synthesized from USP28, batch number M050705. The 0.975 mg / ml solution was further diluted with brine to 0.65 and 0.43 mg / ml solution.

Both compound and vehicle were administered to mice at the body weight of the animals daily treated with a volume injection of 0.1 mL / 10 g body weight. Seven groups of 10 mice per group were injected intravenously (q4d * 4, 13, 17, 21 and 25) with 4 injections every 4 days, as follows: Group 1-Saline , Groups 2, 3 and 4-8.0, 5.3 and 3.5 mg / kg / dose, respectively, with DOX, group 5. 6 and 7-treated with 9.75, 6.5 and 4.3 mg / kg / dose with DOX orotate, respectively ( Based on molecular weight of DOX = 580 and molecular weight of DOX orotate = 708).

And measuring the tumor, and determining the volume by using the L * W ^2/2 = ㎣ formula and and the weight was calculated by assuming a 1㎣ = 1㎎. This study was terminated at 95 days after tumor transplantation.

At day 26 (1 day after the last injection), 5 animals were euthanized from each of groups 1, 2 and 5, and hearts were collected to determine doxorubicin levels.

result:

Tumor weight —administration of DOX at doses of 8.0, 5.3 and 3.5 mg / kg / dose (FIG. 5) and administration of DOX orotate at equivalent doses of 9.75, 6.5 and 4.3 mg / kg / dose. (FIG. 6) Intravenous injection (iv) is very effective in inhibiting the growth of CAKI-1 human kidney tumors subcutaneously xenografted in male NCr-nu / nu mice. There is no apparent difference in tumor growth from administration of DOX orotate compared to DOX, indicating that intravenous administration of DOX orotate and DOX hydrochloride have significant antitumor effects (FIG. 7).

Group pair P value Up to 2 times ^a Day 26 weight ^b 1 to 2 User Element (UE) <0.0001 1 to 3 0.000 <0.0001 1 to 4 0.000 <0.0001 1 to 5 User element <0.0001 1 to 6 0.000 <0.0001 1 to 7 0.000 <0.0001

Example  3. Within the Mouse Heart Doxorubicin  Measure

The concentration of doxorubicin was measured by high performance liquid chromatography (HPLC) mass spectrometry / mass spectrometry. This was an average of 7,398 ng / g cardiac tissue for animals treated intravenously with DOX at a dose of 8.0 mg / kg / dose (group 2) and DOX o at a dose of 9.75 mg / kg / dose (group 5). The average of 5,264 ng / g cardiac tissue for animals treated intravenously with rotate. In other words, the tissue levels of doxorubicin in the cardiac tissues of animals treated with DOX orotate were reduced to 28% compared to the tissue levels in the cardiac tissues of animals treated with DOX. Importantly, cardiac tissue samples were obtained 1 day after the last intravenous injection, and therefore, even after 1 day of intravenous administration of two different forms of DOX, there was a clear difference in DOX levels for DOX orotate compared to DOX hydrochloride. It is indicated that there is a decrease. One skilled in the art can expect that the difference in cardiac DOX levels between the two groups can be greater in the long term. This is because the effect of DOX hydrochloride is partly limited by cumulative dose-related cardiotoxicity, which can usually occur late during the course of treatment or exist for months or years after completion of treatment (therapy). This strongly suggests that drug residues may remain in the tissue after the treatment (therapy) is completed. Thus, there is an absolute need to reduce and / or prevent the drug from accumulating immediately at the end of treatment (therapy), as in subsequent short and long periods in normal or non-infected tissue that does not develop cancer. . The present invention provides methods and compositions for making doxorubicin a DOX orotate derivative, which reduces the heart levels of DOX and thus makes the heart safer from the toxic effects of doxorubicin in the heart in the short and / or long term.

The present invention is not limited in terms of the embodiments described in the embodiments which are intended as a description of one aspect of the invention, and any functionally equivalent methods are within the scope of the invention. As a result, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the appended claims.

Those skilled in the art can recognize or identify using mechanical experiments or any equivalents to certain embodiments of the invention described herein. Such equivalents are intended to be included in the appended claims.

Claims (2)

A compound having the structure of formula (2) for the treatment of primary or metastatic tumors, proliferative hematopoietic disorders or leukemia.
(2)

A compound having the structure of Formula 3 to reduce the level of doxorubicin in cardiac tissue and to prevent cardiomyopathy.
(3)

Images