JPH0457646B2 - - Google Patents

Description

[Detailed description of the invention]

This application was filed on December 15, 1980.
This is a continuation in part of my pending application No. 216197, entitled Treatment of Malignant Tumors with Ribofuranosylthiazole-4-carboxamide and related compounds, and the entire disclosure of which is hereby incorporated by reference. The present invention provides a path to the treatment of malignant tumors in vivo using the compound 2-β-D-ribofuranosylthiazole-4-carboxamide and related derivatives such as esters thereof. The control of malignant tumors in humans and animals remains an unrealized goal. Over the past few decades, remarkable advances have been made in our understanding of malignancy. However, the conquest of malignant disease states has not yet been achieved. Common treatments for both humans and other useful animal species suffering from malignant tumors currently include chemotherapy by surgical removal of the tumor in the affected animal, local radiotherapy, and administration of chemotherapeutic agents to the animal. In many patients with malignant tumors, death is not due to the initial tumor, but instead due to the initial tumor metastasizing to subsequent parts of the host. If the initial tumor is detected early, it can be removed, usually by surgery, radiation or chemotherapy, or a combination thereof. However, these primary tumor metastatic communities are very difficult to detect and eliminate, and their unsuccessful treatment has become a serious medical problem. Tumors are usually classified as either benign or malignant. Malignant tumors are identified as benign by their ability to invade surrounding tissues and to spread to distant sites by metastasis. Some organs are more susceptible to metastasis than others. Included in this group are lungs;
It has a brain, liver, ovaries and adrenal glands. Furthermore, it has been suggested that surgery and radiation directed at the initial tumor may actually promote metastasis in some cases. Given the inability of current cancer therapies to inhibit malignant tumors and their metastasis, it is clear that there is a need for additional chemotherapeutic agents. In a paper entitled Synthesis and Antiviral Activity of Certain Thiazole C-Nucleosides (J.Med.Chem. 1977, Vol. 20, No. 2256), I and my collaborators reported that the compound 2-β-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O-acetyl-β-
Synthesis of D-ribofuranosyl) thiazole-4-carboxamide and in vitro test system of three viruses, hydatid simplex virus type 1, parainfluenza virus type 3, and 3
Preliminary antiviral activity in vitro performed using a type of rhinovirus is described. The in vitro activity of the compound 2-β-D-ribofuranosylthiazole-4-carboxamide against these three viruses was only modest. Compound 2-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)thiazole-4
- Regarding carboxamides, only mild activity was observed with simple vesicle virus type 1, but no activity was observed with parainfluenza type 3 and rhinovirus type 13. Although some marginal in vitro antiviral activity was seen, quite to the contrary, 2
-β-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O
-acetyl-β-D-ribofuranosyl)thiazole-4-carboxyamide The in vivo antiviral test of both was negative as judged by the number of deaths of test animals. In this in vivo study, 2-β-D-ribofuranosylthiazole-4
- The number of test animal deaths using both carboxamide and 2-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)thiazole-4-carboxamide is the same as the number of placebo control animal deaths. Compound 2-β-
D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)thiazole-4
- shows that neither of the carboxamides demonstrated useful in vivo antiviral activity. 2-β-D-ribofuranosylthiazole-4-
For the above in vitro antiviral testing of both carboxamide and 2-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)thiazole-4-carboxamide, these compounds were compared with known antiviral compounds. Ribavirin
(RIBAVIRIN) was tested against viruses with positive antiviral activity. 2-β-D
In terms of the preliminary in vitro peripheral activity of ribofuranosylthiazole-4-carboxamide against these test viruses, the spectrum of activity of 2-β-D-ribofuranosylthiazole-4-carboxamide is similar to that of the compound ribavirin. It seems likely. Ribavirin is known to be an active in vitro and in vivo antiviral agent, and is also known to exhibit no apparent antitumor activity. Furthermore, certain derivatives of ribavirin, such as its 5'-monophosphate, are also known to be inactive as antitumor compounds. 2
The preliminary in vitro antiviral activity of -β-D-ribofuranosylthiazole-4-carboxyamide was compared with that of ribavirin.
It is reasonable to assume that ribofuranosylthiazole-4-carboxamide would exhibit the same positive in vivo antiviral activity and negative antitumor activity as ribavirin. Quite contrary to this, the compound 2-β-D-ribofuranosylthiazole-4-carboxamide had no useful in vivo antiviral activity and, quite unexpectedly, showed positive antitumor activity. . Compounds 2-β-D-ribofuranosylthiazole-4-carboxamide and 2-(2,3,5
-tri-O-acetyl-β-D-ribofuranosyl)thiazole-4-carboxamide and 2
The esters thereof, including -(5-O-phosphoryl-β-D-ribofuranosyl)thiazole-4-carboxyamide, have been found to exhibit significant antitumor activity, making them useful as antitumor agents in vivo. Ta. BRIEF SUMMARY OF THE INVENTION The compound 2-β-D-ribofuranosylthiazole-4-carboxamide was found to exhibit significant antitumor activity in vivo. The present invention relates to the use of this compound and certain related derivatives for the treatment of malignancies in warm-blooded animals. According to the present invention, 2-β-D-ribofuranosylthiazole-4
- The antitumor properties of carboxamides and their related esters extend to warm-blooded animals at least and 0.1% by weight based on the total weight of the composition as active ingredients: (In the formula, R ₁ and R ₂ are H or C ₁ -C ₁₈ acyl, and R ₃ is H, C ₁ -C ₁₈ acyl or

and physiologically compatible salts thereof. A more preferred group of compounds is where R ₁ and R ₂ are H or C ₁ -C ₈ acyl and R ₃ is H, C ₁ -C ₈ acyl or

Compounds of the formula and physiologically compatible salts thereof. Preferred acyl groups for R ₁ , R ₂ and R ₃ include acetyl, propionyl, butyryl, isobutyryl and benzoyl. Compatible salts include alkali metal and ammonium or substituted ammonium salts such as the sodium, potassium and ammonium salts. Preferably, when R ₁ and R ₂ are H, R ₃ is
OH, _C1 - _C8 acyl or

[Formula] and when R ₁ and R ₂ are C ₁ - C ₈ acyl, R ₃ is C ₁ -
Preferably it is a C ₈ acyl. For use in the pharmaceutical compositions of the present invention, a pharmaceutical carrier is utilized, which is selected so that an appropriate concentration of the active ingredient of the present invention can be administered by injection as a solution or suspension to a diseased warm-blooded animal. It is preferable to Depending on the host with the malignant tumor, the type of tumor, and the location of the tumor, administration by injection may be intravenous,
Intramuscular, intracerebral, subcutaneous, or intracavitary injection. Alternatively, the compositions of the invention may be formulated in suitable pharmaceutical carriers for administration by other routes, such as oral, ocular, topical or suppository administration. Detailed Description Compound according to the present invention, compound 2-β-D-ribofuranosylthiazole-4-carboxamide,
is preferably prepared as described in Example 1.
An alternative synthesis of this compound is provided by J.Org.
Chem.), Volume 41, No. 2619764074. 2-β-D-ribofuranosylthiazole-4-
Esters of carboxamides (compound 1), such as 2-(2,3,5-tri-O-acetyl-β
-D-ribofuranosyl)thiazole-4-carboxamide (compound 2) or 2-(5-O-phosphoryl-β-D-ribofuranosyl)thiazole-
4-Carboxamide (Compound 3) is prepared as described in Examples 2 and 3, respectively. Additionally, other esters, such as the monoester 2-
(5-O-acetyl-β-D-ribofuranosyl)
Thiazole-4-carboxamide (compound 4),
is prepared as in the synthesis described in Example 4. Other preferred carboxyesters of the invention are obtained by replacing acetic anhydride with a suitable anhydride such as propionic anhydride, butyric anhydride or benzoic anhydride. Alternatively, suitable acid chlorides can be replaced by acid anhydrides. Esters of Compound 1 serve for delivery of the compound in the affected host. Esters of such compounds include one or two sugar moieties of compound 1.
may be generated by reacting one or more hydroxyl groups with a suitable reversible blocking group that can be cleaved from the parent compound 1 in vivo by some in situ chemical or enzymatic reaction. For reaction with hydroxyl groups, esters such as, but not necessarily limited to, acyl and phosphoryl esters are contemplated. Acyl groups are linear, branched, substituted, unsaturated, saturated or aromatic acids such as, but not necessarily limited to, acetic acid, trifluoroacetic acid, propionic acid, n-butyric acid, isobutyric acid, valeric acid, caproic acid, pelargonic acid, enanthic acid, caprylic acid, lactic acid, acrylic acid, propargyl acid, palmitic acid, benzoic acid, phthalic acid,
It can be selected from the group consisting of salicylic acid, cinnamic acid and naphthoic acid. If a phosphoryl group is selected, a phosphoryl ester can be produced as the free acid or as a salt. Compatible salts of the phosphate moiety of the phosphoryl ester include, but are not necessarily limited to, alkali and alkaline earth salts such as sodium, potassium, calcium, magnesium, lithium, ammonium and substituted ammonium, trialkylammonium, dialkylammonium, alkylammonium, e.g. It may be selected from the group consisting of triethylammonium, trimethylammonium, diethylammonium, octylammonium, cetyltrimethylammonium and cetylpyridium. Preferred forms of the esters of the invention include compounds 2, 3 and 4. In addition to these, other tri-O-acyl esters such as 2',
3',5'-tri-O-benzoyl is mentioned. Furthermore, other monoesters such as 5'-O-benzoyl may also be mentioned. Typically, preferred carboxyesters include _C1 - _C18 acyl.
A more preferred group includes _C1 - _C8 acyl. When phosphoryl esters are utilized, it is preferred to form the phosphate group as a salt, preferably as a sodium salt or other alkali metal salt or ammonium salt. As shown in the examples herein, the ester form of Compound 1 is useful in diseased hosts to deliver the compound to the affected area. As shown in the examples, tri-acetyl ester (compound 2) is shown to be an effective antitumor agent when injected intraperitoneally into affected hosts. The above triacetyl compounds and other acyl esters of any of Compound 1 may be prepared by a suitable enzyme such that the ester can be enzymatically cleaved to Compound 1 in the acidic environment of a biological fluid such as the stomach or in vivo. It is thought that Compound 1 is hydrolyzed in an environment containing I do not wish to be bound by theory, but if we use a phosphoryl ester of compound 1, e.g. I think we will derive compound 1 in situ. As shown in the examples, Compound 3, the phosphoryl ester of Compound 1, is shown to be an effective antitumor agent when injected into a diseased host. It is not known here whether the activity is expressed as the 5'-phosphate or whether it is cleaved off by the enzyme to form compound 1. Furthermore, compound 1 can also proceed to compound 3 by other enzymatic reactions in situ. In any case, both Compound 1 and Compound 3 are found to be effective in vivo antitumor agents as shown by the examples. In the practice of this invention, it is appropriate to mix Compound 1, or the selected ester form thereof, with a suitable pharmaceutical carrier, which may be alone, such as sterile water, or It may also be a complex carrier containing appropriate agents to appropriately mimic a certain biological environment, ie, a PH and salt adjusted solution suitable for intravenous, intramuscular or other injection. Selection of a suitable drug carrier takes into account the type of tumor, the site of the tumor, and the health and age of the host.
Furthermore, when using the ester form of Compound 1, the chemical reactivity of the ester is also taken into account. Thus, it is preferred to suspend or dissolve the carboxyacyl ester in a suitable non-acidic medium. It is also suitable to use phosphoryl esters in the presence of suitable buffers or as salts as described above. Compound 1 or any other compound of the invention,
Preferably, Compound 1 or a derivative thereof is mixed with a suitable pharmaceutical carrier such that the compound is suitably dissolved in the carrier. Alternatively, however, suspensions, emulsions and other formulations of the compounds of the invention can also be used, if indicated. Pharmaceutical carriers, in addition to the solubilizing or suspending agents included therein, also include suitable diluents, buffers, surfactants and other similar agents typically employed in pharmaceutical carriers. However, the overall composition of the drug carrier is
It must be selected in a manner that is compatible with the concentration of active ingredient and other parameters standard in the pharmaceutical industry. Compound 1 or other compounds of the invention are suitably mixed with the pharmaceutical carrier present in a concentration of at least 0.1% by weight of the total composition. Preferably, it is present in the pharmaceutical carrier at a concentration of about 10 to about 90% by weight of the total composition. An effective amount of Compound 1 or other compounds of the invention will typically be applied per day to the total body weight of the warm-blooded animal being treated.
Ranging from approximately 2.5mg to approximately 200mg per kg. This range is from 12.5mg/Kg to 100
Preferably it is mg/Kg. An even more preferred range is approximately 15 mg/Kg to approximately 50 mg/Kg. As with the other factors mentioned above, the amount of compound used to treat an affected animal must take into account parameters such as tumor type, tumor site, compound administration mode, and host body size and condition. .
In any event, the actual amount must be sufficient to provide an adequate chemotherapeutically effective amount of the drug to the host, and is within the skill of the art in determining what is described herein. is easy. In at least one study, compound 1 of the invention was
When injected into tumor-bearing animals at a dose of 2000 mg/Kg, no animals died due to Compound 1 toxicity on the day of the toxicity test. Even in a host diagnosed with an undiagnosed disease caused by a malignant tumor, if there is any possibility of cure in the undiagnosed host, as is commonly done in today's cancer chemotherapy, it is beyond the scope of toxicity. Excess amounts may be indicated. As in the following illustrative example, hosts with tumors are administered the indicated test compound once daily. Depending on the clinical situation, the daily dosage of Compound 1 or any other compound of the invention may be given as in the example; however, the daily dosage may be divided into several unit doses and the total daily dosage It can also be given in dosages. Thus, for example, at the 50 mg/Kg dosage level, patients would receive 12.5
Doses of mg/Kg may be given. Compositions used to inhibit malignant tumors in warm-blooded animals are prepared by adding Compound 1 or any other compound of the invention to a pharmacologically compatible solvent, followed by sterilization and appropriate encapsulation. Suitably, the preparation is carried out by filling known concentrations into vials to be obtained. The appropriate dose of compound is then removed from the vial and administered to the host by injection. Example 1 2-β-D-ribofuranosylthiazole-4-
Carboxamide, Compound 1 Ethyl 2-(2,3,5-tri-O-benzoyl-β-D-ribofuranosyl)-thiazole-4
- Carboxamides are described by Srivastova et al., J.D., herein incorporated by reference. Med. Chem. (J.Med.Chem.), 1977, Volume 20, No.
Use as made in No. 2256. Ethyl 2-(2,3,5-tri-O-
Benzyl-β-D-ribofuranosyl)thiazole-4-carboxamide (5.0 g, 8.31 mmol)
A concentrated solution of is stirred with methanolic ammonia (saturated at 0° C., 100 ml) in a pressure bottle at room temperature for 2 days. The solvent was evaporated and the residue was placed on a column of silica gel (100 g) packed in ethyl acetate (2.5 x 35
chromatography through cm). Solvent system (ethyl acetate-1-propanol-water, 4:
When the column is eluted at a ratio of 1:2 (V/V; top layer), methyl benzoate and benzamide move quickly. Most slowly moving UV and sugar-
The sugar-positive fractions are collected and the solvent is evaporated under reduced pressure. The residue thus obtained (syrup-like) was easily crystallized from ethanol-ethyl acetate to yield 1.6 g (74%) of pure product,
Compound 1: mp144−145°C; [α] ²⁵ _p −14.3° (C1,
DMF); UV〓 _nax PH ¹ 237nm (8640); UV〓 _nax PH
¹¹ 238nm (8100); ¹ HNMR (Mc ₂ SO−d ₆ ) δ7.5−
7.8 [S(br), 2, CONH ₂ ] ¹ HNMR(Mc ₂ SO−d ₆
−D ₂ O) δ4.99 (d, 1, J−5H ₂ , H ₁ ′), 8.25
(S, 1, H ₅ ), analysis (C ₉ H ₁₂ N ₂ O ₅ S) C, H,
Provide N, S, Example 2 2-(2,3,5-tri-O-acetyl-β-
D-ribofuranosyl)thiazole-4-carboxamide, compound 2 Acetic anhydride (2.0ml) was added to anhydrous pyridine (16ml).
of Compound 1 (1.04 g, 4 mmol) in an ice-cold solution and the reaction solution was stirred at room temperature for 17 hours. The solvent is evaporated under reduced pressure, the residue is dissolved in ethyl acetate, the solution is washed with water and dried (MgSO ₄ ). The ethyl acetate portion was evaporated under reduced pressure;
The residue thus obtained was crystallized from water to yield 1.4
g (90%) of compound 2 as white needles;
mp103°C; ¹ HNMR ( _CDCl3 ) 2.1 (3S, 9, tri-O-acetyl), 6.2 and 7.15 [S(br) pair,
2, CONH ₂ ], 8.2 (S, 1, H ₅ ). analysis,
(C ₁₅ H ₁₈ N ₂ O ₈ S) C, H, N, S. Example 3 2-(5-O-phosphoryl-β-D-ribofuranosyl)thiazole-4-carboxamide (2-β-D-ribofuranosylthiazole-4
-carboxamide 5'-phosphate), Compound 3 Water (151 mg, 8.4 mmol) was carefully combined with freshly distilled phosphoryl chloride (2.0 g, 13.2 mmol), pyridine (1.21 g, 14.4 mmol) and acetonitrile (2.3 g, 56.7 mmol). mmol) (kept at 0° C. with stirring).
2-β-D-ribofuranosylthiazole-4-carboxamide (compound 1) (800 mg, 3.0 mmol, dried over P ₂ O ₅ and powdered) was added to this solution and the reaction mixture continued. and 0℃ for 4 hours
Stir at . Pour the reaction mixture into ice water (approximately 50 ml)
Pour into the solution and adjust the pH to 2.0 with 2N sodium hydroxide. This solution is applied to a column of activated carbon (20 g) and the column is thoroughly washed with water until the eluate is free of salt. The column is eluted with an ethanol-water-concentrated ammonium hydroxide (10:10:1) solution and fractions (25 ml each) are collected. Purification (tlc,
Silica gel, acetonitrile-0.1N ammonium chloride (7:3)) Fractions containing the nucleotide (compound 3) are collected and evaporated to dryness under reduced pressure.
Dissolve the anhydrous residue in water, dowex 50W−X8 (20−50 mesh, H ⁺ type, 15
ml) through the column. Wash the column with water and collect the fractions containing the nucleotides. Concentrate the solution to a small volume (5 ml) and use a Dowex 50W-
Pass through a column of X8 (20−50 mesh, Na ⁺ form, 15 ml). Wash this column with water. The fraction containing the nucleotides is lyophilized. The residue is triturated with ethanol, collected by filtration and dried (P ₂ O ₅ ) to yield 560 mg (47%) of compound 3 in crystalline form as monosodium dihydrate. Analysis, Calculated as C ₉ H ₁₂ N ₂ O ₈ PSNa・2H ₂ O: C, 27.13; H, 4.04; N,
7.04; P, 7.78; S, 8.05. Actual values: C, 27.42; H, 3.87; N, 7.07; P, 8.03; S, 8.41. Example 4 2-(5-O-acetyl-β-D-ribofuranosyl)thiazole-4-carboxamide, compound 4 2-(2,3-O-
Isopropylidene-β-D-ribofuranosyl)thiazole-4-carboxamide (1.5 g, 5 mmol) (Fuertes et al., J.Org.Chem., Vol. 41, No. 26 issue,
1976, prepared as 4074) is cooled on an ice-water bath and acetic anhydride (2.5 ml) is added with gentle stirring. Warm the reaction solution to room temperature,
Continue stirring for 15 hours. Evaporate the solvent under reduced pressure,
The residue is dissolved in ethyl acetate and washed with water.
The ethyl acetate portion was evaporated under reduced pressure and the residue was reduced to 80%
Dissolve in % acetic acid (25 ml). The solution is heated on a steam bath for 30 minutes and the solvent is evaporated under reduced pressure. The residue is dissolved in ethyl acetate, washed once with water and dried over MgSO ₄ . The ethyl acetate portion was evaporated and the crude product was dissolved in silica gel (100 g,
(packed in chloroform) column and eluted with 20% (v/v) ethyl acetate in chloroform. Fractions containing nucleotides are pooled and evaporated to yield 1.05 g (70%) of compound 4.
_{( C11H14N2O6S ) .} Examples 5 to 12 below are given as demonstrations of Compound 1 and other illustrative compounds of the invention. In these examples, the efficacy of the compounds is demonstrated using standard tests against certain malignancies. The tests used in these demonstrations were conducted under the guidance of the International Cancer Research Institute, Division of Cancer Therapy, and Developmental Therapeutic Project.
This study was conducted using those standard protocols and methods and was supervised by this agency. All tests were conducted in accordance with these agreements, and all studies were evaluated based on the criteria specified by these agreements. The following representative examples illustrate the robust activity exhibited by illustrative compounds of the invention in the International Cancer Research Institute's Screening Tumor Format. In the examples below, the abbreviation IP stands for intraperitoneal;
Also, IV stands for intravenous. Mean and median survival times are calculated by the International Institute for Cancer Research, Division of Cancer Therapy, Developmental Treatment Plans, Screening Data List Professor 14 (Revised 6/78). The contents of the Quantitative Screening Data List, including appropriate revisions, are included herein by reference. In the following demonstration example, excipients used as carriers for drugs were injected into control animals at the same level as the excipients used in drug-treated animals to eliminate the effects of any excipients in this study. (excluding any drugs therein). Example 5 As an indicator of regenerative activity, compound 1 of the invention is screened against L-1210 lymphocytic leukemia in vivo using CD ₂ F ₁ male mice as the test species. The selected efficacy parameters are based on the median survival time of drug-treated animals versus appropriate control group animals. Both drug-treated and control animals are inoculated IP with 10 ⁵ seed cells of L-1210 lymphoid leukemia into the ascites fluid. One day after tumor inoculation, animals in drug groups are placed under treatment regimen of Compound 1 at dosage levels as listed in Table 1 below. Animals in drug-treated groups are inoculated by IP injection with the test compound appropriately diluted in water at the stated dosage once daily for 5 days. Day 6 is selected as the indicator of drug toxicity. In this example, all drug-treated animals survived for 6 days. Death of drug-treated animals after 6 days is therefore
Death was due to tumor and not drug toxicity. The median date of death in the control group was 8.5 days. As shown in Table 1 below, the median date of death in the drug-treated group was longer than that in the control group at all levels of the treatment drug, and was clearly longer at 50 mg/Kg (drug dose/body weight of test animal) and above. It was. 1st below
The results shown in the table indicate that the drug exhibited positive activity in this fold assay. % of drug-treated animals/control animals greater than or equal to 125% is considered positive drug activity.

[Table] Example 6 Compound 2,2-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)thiazole-4
- Carboxamides are screened in the same manner as described in Example 5, but the tumor line used as the test tumor is p388 lymphocytic leukemia. ¹⁰⁶ seed cells are used to generate tumors in both control and drug-treated animals. Use the same strain of mice, but use female mice instead of males. The test results are based on the mean survival time and are expressed as T/C percentage (treated animals/control animals) as per Example 5. For drug-treated animals, treatment begins one day after tumor inoculation and drugs are given at the dosage levels listed in Table 2 below. Continue drug treatment for 9 days,
Drug toxicity is determined on day 6 as in Example 5.
At the 100 mg/Kg level, only one animal failed to survive beyond the toxicity cut-off date. The mean day of death in the control group was 10.2 days, while treated animals survived for more than 15 days even at the lowest level of drug treatment. As in Example 5, a 125% increase in longevity in treated animals compared to control animals is considered an indicator of a positive drug response.

[Table] Compound 1 is also similar to Example 6a, b and c.
Compound 2, 2-(2,3,5-
tri-O-acetyl-β-D-ribofuranosyl)
Thiazole-4-carboxamide is further prepared in Example 7
It was shown to be active against P388 lymphocytic leukemia. In both of these examples, the compounds successfully passed the DN2 (Decision Net) criteria of the International Institute for Cancer Research testing. Examples 7 and 8
Here, _{CD2F1 female} mice were challenged with P388 lymphocytic leukemia tumors. Median survival times of drug-treated animals are compared to appropriate control animals, and both test compounds are considered active anti-tumor agents based on this criterion. The test period was 30 days in both Examples 7 and 8. In Examples 7 and 8, as in Examples 9 and 10 below, any animal in the drug-treated group that survives beyond the end of the study period is then evaluated and placed in one of the three groups. The first group was called cured, meaning the animals were successfully treated for the tumor. The second group is named no-takes, meaning that the survival of the animals is considered to be due to failure of tumor engraftment. The remaining group is referred to as tumor survivors, meaning that the animals survived beyond the study termination date but cannot be classified as either cured or non-receiving. Both Examples 7 and 8 used 30 animals as a control group and drug treatment groups with 6 animals each for each dose level shown in Tables 3 and 4 below. In both Examples 7 and 8, tumors in both control and drug-treated groups were induced by IP inoculation with tumor seed cells on day 0;
Drug treatment is then started on the first day. Both Examples 7a and 8 use saline-tween/80 as the drug excipient. example
In 7b and 7c, water is used as the drug excipient. For both the control and drug-treated groups in Examples 7 and 8, test animals are inoculated IP with 10 ⁶ seed cells of P388 lymphocytic leukemia on day 0. Examples 7 and 8
In both drug groups, treatment begins on day 1, with drug administered IP once daily for 9 days. Day 6 is the censoring date for death due to drug toxicity. In only one case, example 7b,
Animal deaths due to drug toxicity were observed. Treatment efficacy is determined by comparing the median survival time of drug-treated animals to the median survival time of control animals;
Expressed as a percentage increase in treated animals/control animals (T/C) as in Example 5. Example 7a In this example, drug-treated animals are given IP injections at the drug levels used in Table 3 below. Six animals are treated at each dose level. None of the control animals survived for more than 18 days, and the median date of death was
It was 12.6 days. Median dates of death for drug-treated animals are shown in Table 3a below. At the 50 mg/Kg level, one drug-treated animal survived;
This animal was marked as not received. Example 7b This example is conducted as in Example 7a at the dosage levels as shown in Table 3b below. Surviving animals at both the 700 and 800 mg/Kg levels were considered cured.
None of the control animals survived for more than 12 days, and the average date of death for the control group was 11 days. Example 7c This example proceeds as in Example 7a above at the dosage levels shown in Table 3c below. All control animals died by day 14, with an average date of death of 11.9 days. At the 500 mg/Kg level, one animal was deemed cured. Example 8 Compound 2, 2-(2,3,5-tri-O-
Acetyl-β-D-ribofuranosyl)thiazole-4-carboxamide is tested as in Example 7a at the dosage levels shown in Table 4 below. None of the controls survived more than 18 days, and the average date of death was
It was 12.6 days. At the 50 mg/Kg level, one animal that survived was declared a non-receiver. Both compounds 1 and 2 are found to be active antitumor agents in the fold studies presented in Examples 7 and 8.

【table】

【table】

【table】

Table: Compound 1 was shown to be active against L-1210 lymphoid leukocytes as in Example 9 and successfully passed the DN2 criteria of the International Cancer Research Institute test.
In Examples 9a and 9b, _{CD2R1 male} mice were used to
I tried -1210 lymphoid leukocytes. The mean survival time of the test animals was compared to appropriate control animals, and based on this criterion, Compound 1 was considered an active anti-tumor agent. The test period was 30 days. The test results are expressed as T/C as in Example 5. Example 9a uses 24 control animals and the following 5a
Six animals were used for each drug dose as indicated in the table. In Example 9b, 40 control animals were used and 10 test animals were used for each drug dose level as shown in Table 5b below. In both the control group and the drug test group, tumors were injected with tumor seed cells on day 0.
Challenge by IP inoculation, then start drug treatment on day 1. Example 9a uses water as the drug excipient and Example 9b uses Saline as the drug excipient. In Examples 9a and 9b, test animals are inoculated IP with 10 ⁶ seed cells of L-1210 lymphoid leukocytes on day 0 for both control and drug-treated groups. In example 9a,
Drug treatment started on day 1, with Compound 1 once daily 9
Administer for days. Day 5 is the censoring date for death due to drug toxicity. In case 9b, only one patient died due to drug toxicity. Treatment efficacy is determined by comparison of the mean survival time of drug-treated animals to the mean survival time of control animals and is expressed as a percentage increase in treated animals/control animals (T/C) as in Example 5. Example 9a In this example, drug-treated animals are injected IP at dose levels as shown in Table 5a below. Six animals are used for each dose level. In control animals
None of the animals survived for more than 10 days, and the average date of death was 9.7 days. The average date of death for drug-treated animals is as shown in Table 5a below. Example 9b In this example, drug-treated animals are injected IP at the dose levels shown in Table 5b below. Ten animals are treated at each dose level. None of the control animals survived for more than 10 days, and the average date of death was 9.0 days. The average date of death for treated animals is shown in Table 5b below.

【table】

[Table] Compound 1 against Lewis liver cancer in case 10
It was shown to be as active as in the US and successfully passed the DN2 criteria of the International Cancer Research Institute test. example
_In 10 _{, B6D2F1} male mice were used and challenged with Lewis liver carcinoma. The median survival time of test animals is compared to appropriate control animals, and based on this criterion Compound 1 is considered an effective anti-tumor agent. In Example 10, 40 animals are used for the control group and 10 test animals are used for each of the dose levels shown in Table 6 below. For both control and drug-treated groups, tumors are induced by IV injection on day 0, followed by drug treatment beginning on day 1. Example 10 uses water as the drug excipient. In Example 10, animals are inoculated with a homogenate of 10 ⁶ seed cells of Lewis hepatoma on day 0 for both control and drug-treated groups. In Example 10, drug treatment was
Starting on day 1, Compound 1 is administered once daily for 9 days. Day 5 is the censoring date for deaths due to drug toxicity. In this case, there were no deaths due to drug toxicity. Treatment efficacy is determined by comparing the median survival time of drug-treated animals to the median survival time of control animals, which is expressed as a percentage increase in treated animals/control animals (T/C) as in Example 5. Expressed as The test period is 60 days, and at the end of the 60 day period, surviving animals in the test group are evaluated as cured, nonrecipient, or tumor survivors as in Example 5 above. Example 10 In this example, drug-treated animals are injected IP with the dose levels shown in Table 6 below. For each dose level
Use 10 animals. None of the control animals survived for more than 23 days, and the median date of death was 18.4 days. At test levels of 400, 200 and 25 mg/Kg, all animals survived the 60 day test period. Due to this fact, the T/C shown in Table 6 below
The ratio is constant when calculated from the median survival date of 60 days for treated animals and the median death date of 18.4 days for control animals. In Example 10, all 10 surviving test animals at both the 200 and 400 mg/Kg levels were determined to be cured.
At the 100 mg/Kg level, 8 animals were cured, 1 tumor survivor and 1 animal died on day 46. 50mg/Kg
At the level, 9 were cured and 1 died on day 47. Compound 1 was shown in Example 10 and was found to be an active anti-tumor agent in a fold study.

[Table] As shown in Example 10 above, Compound 1 shows significant activity against Lewis liver cancer. Lewis liver cancer is a prominent example of a metastatic tumor system. The test and control animals of Example 10 are inoculated IV with tumor homogenate. The dramatic features of this tumor appear in the liver. As mentioned above, the ability to metastasize is the only characteristic that distinguishes malignant tumors from benign tumors. In Example 10, not only was the median survival time of the drug-treated animals surprisingly prolonged, but at the end of the study period there was at least 80% cure at all but one dose level, and at two of those levels. There was 100% cure. Example 11 Compound 3, 2-(5-O-phosphoryl-β
-D-ribofuranosyl)thiazole-4-carboxamide was shown to be active against L-1210 lymphoid leukocytes as in Examples 11a and 11b. These examples are conducted essentially the same as Example 9 above unless otherwise noted. Compound test dose levels are as follows in Section 7a in each of Examples 11a and 11b.
and as described in Table 7b. In these examples, 36 control animals were used and 6 test animals were used for each drug dose level as shown in Tables 7a and 7b. Saline was used as a drug excipient. No drug toxicity was observed in the test animals in either Example 11a or 11b. None of the control animals survived longer than 10 days in Example 11a, with an average date of death of 8.3 days, and in Example 11b, none survived longer than 11 days, with an average date of death of 10.1 days. . In both the control group and the drug test group, tumors are induced by IP inoculation of tumor seed cells on day 0, and then drug administration begins on day 1, with Compound 3 administered once daily for 5 days. The test results are expressed as T/C as in Example 5.

【table】

EXAMPLE 12 In Example 12, Compound 1 is administered IP to a group of AKD ₂ F ₁ mice that are infected and develop brain tumors by intracranial inoculation with Lewis liver seed cells. As can be seen from the results in Table 8 below, the effects of Compound 1 on affected animals
IP inoculation resulted in a reduction in brain tumors, indicating that IP inoculation of Compound 1 to affected animals successfully crossed the blood-brain barrier. Thirty-two control animals were used in this study, and none of the control animals survived for more than 11 days, and the average date of death for the control group was 9.6 days. Eight test animals were used for each drug dose level except for the 300 mg/Kg level as shown in Table 8. Water was excluded as a drug excipient. In both the control and test groups, tumors were induced on day 0, and drug treatment with Compound 1 administered once daily for 9 days was started on day 1.
The test results are expressed as T/C as in Example 5.

Table: In many brain disease states, both pathogenic and host dysfunction, treatment is inhibited because the drug does not cross the blood-brain barrier. In certain cases where appropriate treatments for disease states are known, effective concentrations of appropriate chemotherapeutic agents do not cross the blood-brain barrier when they are intracranial during the treatment of these diseases. can cause complications. As can be seen from Table 8, the indication that Compound 1 successfully crosses the blood-brain barrier is very promising for the treatment of brain tumors. The following Representative Examples 13-17 illustrate formulations containing active compounds of the invention in illustrative pharmaceutical compositions using illustrative carriers. Among these examples, Example 13 illustrates the use of a compound of the invention in a host animal as an injectable solution suitable for intravenous or other forms of injection. Example 14 describes an oral syrup formulation, Example 15 describes an oral capsule formulation, and Example 16 describes an oral tablet formulation. Example 17 describes the use of a compound of the invention as a suitable suppository. Examples 13-17 list the ingredients and then describe the method of making the compositions. Example 13 Injection Example 13a Compound 1 Compound 1 250mg-1000mg Water for Injection USP appropriate amount Dissolve Compound 1 in water and pass through a 0.22μ filter. Add the filtrate to an ampoule or vial, seal, and sterilize. Example 13b Compound 3 Compound 3 as the sodium salt 25 mg - 1000 mg Water for Injection USP dosage Prepared as in Example 3a above. Example 14 Syrup Example 14a Compound 1 250 mg active ingredient/5 ml syrup Compound 1 50 g Purified water USP 200 ml Thierry syrup as appropriate or 1000 ml Dissolve Compound 1 in water and add the syrup to this solution with gentle stirring. Example 14b Compound 3 250mg active ingredient/5ml Compound 3 as syrup sodium salt 50.0g Purified water USP as required or 200ml Thiery syrup as required or 1000ml Example 15 Capsule Example 15a Compound 1 100mg, 250mg or 500mg Compound 1 500g Lactose USP, anhydrous Appropriate amount or 200g Stelotex powder HM 5g Compound 1 and lactose intensifier
The mixture is combined in a blender consisting of a pair of containers (bar). Blend vigorously for 2 minutes, then blend for 1 minute using the intensifier wand, then vigorously blend again for 1 minute. A portion of the formulation is then mixed with the Sterotex powder, passed through a #30 film, and passed back into the rest of the original formulation. The mixed ingredients are then blended for 1 minute;
Blend for 30 seconds with an intensifier rod and mix vigorously for an additional minute. 100mg in an appropriately sized capsule,
Capsules containing 250 mg and 500 mg are filled with 141 mg, 352.5 mg or 705 mg of the formulation, respectively. Example 15b Compound 2 100 mg, 250 mg or 500 mg Compound 2 500 g Lactose USP, anhydrous or 200 g Stelotux powder HM 5 g Mix and fill as in Example 15a. Example 15c Compound 4 100 mg, 250 mg or 500 mg Compound 4 500 g Lactose USP, anhydrous or 200 g Stelotux powder HM 5 g Mix and fill as in Example 15a. Example 16 Tablet Example 16a Compound 1 100mg, 200mg or 500mg Compound 1 500g Corn starch NF 200.0g Cellulose, microcrystalline 46.0g Stelotex powder HM 4.0g Purified water appropriate amount or 300.0ml Corn starch, cellulose and Compound 1 mixed in a planetary Combine together in the machine and mix for 2 minutes. Add water to this combination and mix for 1 minute. The resulting mixture is spread on a tray and dried in a hot air oven at 50°C until the humidity reaches 1 to 2%. The dry mixture is then pulverized in a Fitzmill and passed through #RH2B film on medium speed. The Sterotex powder is added to a portion of the mixture and passed through a #30 sieve, which is returned to the original powdered mixture, and the whole is blended by rotating the cylinder for 5 minutes.
Compressed tablets of the total mixture of 150 mg, 375 mg and 750 mg respectively are formed with appropriate size punches as tablets containing 100 mg, 250 mg or 500 mg. Example 17 Suppository Example 17a Compound 1 250mg, 500mg or 1000mg/3g Compound 1 250mg 500mg 1000mg Polyethylene glycol 1540
1925mg 1750mg 1400mg Polyethylene glycol 8000
825mg 750mg 600mg Polyethylene glycol 1540 and polyethylene glycol 8000 are melted together at 60°C and compound 1 is dissolved into this melt. Place this entire mixture into a mold at 25°C to make a suitable suppository. Example 17b Compound 2 250, 500, 1000mg/3g Compound 2 250mg 500mg 1000mg Polyethylene glycol 1540
1925mg 1750mg 1400mg Polyethylene glycol 8000
825mg 750mg 600mg Prepared similarly to Example 17a above.

[Claims]

1. Present in a therapeutically effective amount sufficient to suppress or alleviate symptoms of atherosclerosis26
- A pharmaceutical preparation for suppressing or alleviating symptoms of atherosclerosis, characterized in that it contains hydroxycholesterol as an active ingredient.

Claims (1)

The composition according to claim 1, which is a derivative of the compound. 4. The composition of claim 3, wherein the acyl ester is an acetate ester. 5. The composition of claim 3, wherein the acyl ester is a benzoic acid ester. 6. The composition of claim 1, wherein said compound is a phosphoryl ester of 2-β-D-ribofuranosylthiazole-4-carboxamide. 7. The composition of claim 1, wherein said compound is 2-β-D-ribofuranosylthiazole-4-carboxyamido-5'-phosphate.