IE58434B1

IE58434B1 - Inhibition of tumor development

Info

Publication number: IE58434B1
Application number: IE159785A
Authority: IE
Original assignee: Colgate Palmolive Co
Priority date: 1984-06-27
Filing date: 1985-06-26
Publication date: 1993-09-22
Also published as: LU85976A1; JPH0623108B2; CH668361A5; JPS6140217A; IE851597L; IT8548226A0; MC1672A1; KR880002266B1; FR2566665B1; DE3523263A1; AU4418785A; PT80707B; PT80707A; FI84697B; IL75518A0; WO1986000225A1; DK168191B1; BE902732A; FI852504L; FI84697C

Abstract

Non-toxic-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid when administered orally or systematically inhibits tumor development of tumor cells in vitro and actual tumor development in vitro in warm blooded animals.

Description

This invention relates to the inhibition of development of tumour cells in vitro and actual tumour development in vivo in warm blooded animals.

The disease of cancer results from development of malignant tumours. A vast amount of medical research has been committed to reducing and overcoming the scourge of cancer. To date, a cure for cancer has not been found. However, much has been learned of the mechanism by which warm blooded animals avoid affliction with cancer. The present invention builds on this knowledge to provide a material which inhibits tumour development and a method for inhibiting such development.

Among the cells contained in mammalian body fluids are lymphocytes, monocytes, macrophages and polymorphonuclear cells. These cells act as a natural surveillance system against tumour development in lower mammals, such as rodents, up to humans. In recent years, it has been observed that a particular subpopulation of lymphocytes or lymphoid cells, termed Natural Killer or NK cells, destroy tumour cells and thus prevent development of cancer. The weight of evidence suggests that NK cells possess cytolytic activity related to generation of an active oxygen species such as hydrogen peroxide (H2O2) or oxygen-containing radicals, e.g. hydroxyl anion (·ΟΗ) and superoxide anion (O2·)· The NK cells and active oxygen phenomena are described by Herberman et al, Science, Vol. 214, 2 October 1981, pages 24-30, Roder et al, Nature, Vol. 298, 5 August 1982, pages 569-572 - 2 Nathan et al, Journal of Immunology, Vol. 129, No. 5, November, 1982, pages 2164-2171; and Havier et al, Journal of Immunology, Vol. 132, No. 4, April 1984, pages 1980-1986.

. Of course, there are many compounds which release active oxygen species. However, that factor alone has not meant that such compound could be introduced into a body to supplement the function of NK cells or where tumour formation is not sufficiently . occurring to provide the function of NK cells and inhibit tumour development. Compounds which release active oxygen species generally do so quickly, while effectiveness against tumour development in warm blooded animals such as humans would appear to . require at least a slower and more sustained release rate. Until this invention, this had not been effectively achieved. When oxygen release is too fast both tumourous and normal cells may be attacked.

In U.S. Patent Specification No. 4,041,149, granted August 9, . 1977, to the inventor of the present invention and coinventors. a composition was described in various forms, including as a dental tablet, which inhibited formation of mouth odour in which the active ingredient was a peroxydiphosphate. Peroxydiphosphate . compound differs from most oxygen providing compounds in that it does not provide an initial burst of hydrogen peroxide. Rather, it releases hydrogen peroxide slowly such that when equivalent concentrations are compared to hydrogen peroxide, the amount . of oxygen released by the peroxydiphosphate is one3 10 . . . . tenth the amount of available oxygen released by hydrogen peroxide. Moreover, only about 50% of the active oxygen is released in 20 hours at 25 °C in the presence of alkaline phosphatase or acid phosphatase each of which is present in the bodies of warm blooded animals, including mice, rats, humans, etc.

By means of the present invention, tumour cell development may be inhibited in vitro and actual malignant tumour development in vivo in warm blooded animals, such as rodents ranging up to humans, may also be inhibited.

The present invention provides a method for inhibiting tumour formation by introducing a slow oxygen releasing material into a living host.

In accordance with a first aspect, this invention comprises the use of a peroxydiphosphoric acid or a pharmaceutically acceptable compound derivative thereof in the preparation of an anti cancer agent.

From 0.1% to 10% (by weight) of the composition may be active compound, which may be dissolved or dispensed in the carrier. The composition may be sterile.

The invention can be used in a method of inhibiting the formation, or proliferation of malignant tumour cells, in which a composition comprising a non-toxic dosage amount of from 0.1 to 6 gm per kg body weight of a warm blooded animal of a non-toxic water-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid, or . - 4 the acid itself, in association with, e.g. dissolved or dispersed within, a pharmaceutically acceptable carrier is administered to a warm blooded animal host by oral ingestion in a regimen which provides from . 0.1 to 6 gm per kg body weight of the warm blooded animal per day.

The invention may be used in a method of inhibiting tumour formation in which a composition comprising a non-toxic dosage amount of from 0.1 to 2 . gm per kg body weight of a warm blooded animal having malignant tumour cells of a non-toxic water-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid, or the acid itself, dissolved or dispersed in a pharmaceutical carrier is . administered systemically, for example parenterally, to a warm blooded animal host in a regimen which provides from 0.1 to 2 gm per kg body weight of the warm blooded animal.

The peroxydiphosphate compound (PDP) is in the . form of a non-toxic pharmaceutically acceptable compound, which goes beyond salt indicated in earlier mentioned US. Patent Specification No. 4,041,149. Compounds include alkali metal (e.g. lithium, sodium and potassium) alkaline earth metal (e.g. magnesium, calcium and . strontium), zinc and tin salts as well as organic peroxydiphosphate alkyl, adenylyl, guanylyl, cytosylyl and thymylyl esters and also quaternary ammonium and the like salts. Alkali metal, parti30 . - 5 cularly potassium salt is preferred from among the inorganic cations. The tetrapotassium peroxydiphosphate is a stable, odourless, finely divided, free-flowing, white non-hygroscopic crystalline solid . having a molecular weight of 346.35 and an active oxygen content of 4.6%. Tetrapotassium peroxydiphosphate is 47-51% water-soluble at 0°-61°C, but insoluble in common solvents such as acetonitrile, alcohols, ethers, ketones, dimethyl forraamide, . dimethyl sulphoxide, and the like. A 2% aqueous solution has a pH of about 9.6 and a saturated solution thereof a pH of about 10.9. A 10% solution in water at 25 °C showed no measured active oxygen loss after four months, and at 50JC a 10% solution . showed an active oxygen loss of 3% in 6 months.

The organic salts can be particularly suitable for administration against malignant tumours- From among the organic esters those providing hydrophobic properties such as C^_^2 alkyl radical and those . which facilitate the rapid uptake of peroxydiphosphate moiety by the cells, such as adenylyl, guanylyl, cytosylyl and thymylyl esters, are preferred.

Pharmaceutical carriers suitable for oral ingestion are coated tablets composed of material . which resists breakdown by gastric acids in the stomach pH (about 1-3) since peroxydiphosphate would be inactivated by such gastric acids. Rather, the carriers, with tableted granules of the peroxydiphos phoric acid salt solid material therein, are . dissolved by intestinal fluids which have a higher pH (about 5.5-10) and do not inactivate the peroxydiphos phate, leaving it subject to enzymatic action by phosphatase present in humans or other warm blooded animals. A desirable tablet coating solution is . composed of a fatty acid ester such as N-butyl stearate (typically about 40-50, preferably about 45 parts by weight), wax such as carnuba wax (typically about 15-25, preferably about 20 parts by weight), fatty acid such as stearic acid (typically about . 20-30 parts, preferably 25 parts by weight) and cellulose ester, such as cellulose acetate phthalate (typically about 5-15, preferably about 10 parts by weight) and organic solvent (typically about 400-900 parts). Other desirable coating materials include . shellac and copolymers of maleic anhydride and ethylenic compounds such as polyvinyl methyl ether. Such coatings are distinct from tablets which are broken down in the oral cavity in which the tablet material typically contains about 80-90 parts by . weight of mannitol and about 30-40 parts by weight of magnesium stearate.

Tabletted granules of the peroxydiphosphate salt are formed by blending about 30-50 parts by weight of the peroxydiphosphate salt with about 45-65 parts by . weight of a polyhydroxy sugar solid such as mannitol and wetting with about 20-35 parts by weight of a polyhydroxy sugar compound solution such as sorbitol, screening to size, blending with about 2-35 parts by weight of a binding agent such as magnesium stearate . and compressing the granules into tablets with a tablet compressing machine. The tabletted granules are coated by spraying a foam of a solution of the coating material thereon and drying to remove solvent. Such tablets differ from dental tablets which are typically compressed granules without a special protective coating.

An effective dosage of administration of peroxydiphosphate with a prescribed regimen, when administration is by oral ingestion, is about from 0.1 to 6 g per kg of body weight daily; when administration is systemic, such as by intramuscular, intraperitoneal or intravenous injection, the dosage is about from 0.1 to 2 g per kg of body weight daily.

The invention also extends to a suppository comprising peroxydiphosphoric acid, or a pharmaceutically acceptable compound derivative thereof, and a rectally or vaginally acceptable carrier.

The invention further extends to a pharmaceutical formulation containing peroxydiphosphoric acid, or a pharmaceutically acceptable compound derivative thereof, and a pharmaceutically acceptable carrier, the formulation being other than in the form of a dentifrice, a mouthwash, chewing gum, a lozenge or a sorbitol-containing composition. The invention in addition extends to a pharmaceutical formulation containing peroxydiphosphoric acid, or a pharmaceutically acceptable compound derivative thereof, and a pharmaceutically acceptable carrier, the formulation being substantially free of humectants, ethanol and saccharides.

Physiologically acceptable pyrogen-free solvents are suitable carriers for use in the art-recognized manner for systemic or parenteral administration. Saline solution buffered with phosphate to a physiological pH of about 7 to 7.4 is the preferred carrier for systemic administration. Such solvents are distinct from waterhumectant vehicles typically used in dentifrices. Such solution is typically prepared by sterilizing deionized distilled water, checking to ensure non-pyrogenicity using the Limulus amoebocyte lysate (LAL) test described by Tsuji et al. in Pharmaceutical Manufacturing, October, 1984, pages 35-41, and then adding thereto a phosphate buffer (pH e.g. about 8.5-10) made in pyrogen free sterile water and about from 1 to 100 mgs peroxydiphosphate compound derivative and sodium chloride to a concentration of about 0.5-1.5% by weight. The solution can be - 8 packed in vials for use after being resterilized by passing through a micropore filter. As alternatives, other solutions such as Ringer's solution containing 0.86% by weight sodium chloride, 0.03% by weight . potassium chloride and 0.033% by weight calcium chloride may be used.

Peroxydiphosphate compound (PDP) release hydrogen peroxide slowly in the presence of phosphatase enzymes in accordance with the following . equation: O O 0 II phosphatases il H2O -^2θ2+Ρθ4_3' X4-O-P-O-O-P-O-> -O-O-P-O- -> . owherein X is a non-toxic pharmaceutically acceptable cation or completes an organic ester moiety. Phos20. phatase to break down the peroxydiphosphate is present in saliva as well as in plasma, intestinal fluids and white blood cells. The slow oxygen release is particularly effective in supplementing the effectiveness of NK cells against malignant . tumour cells which respond to peroxydiphosphate therapy. When warm blooded animals are treated with PDP in accordance with the present invention it is desirable to provide a regimen whereby treatment continues at least until tumours are regressed.

. The following examples illustrate the invention.

All amounts are by weight unless otherwise indicated. - 9 EXAMPLES Examples IA and IB: In Vitro Study of PDP Tumour Cytotoxicity.

In this study the effects of PDP are examined at . different concentrations on the growth of murine myeloma (SPg line) cells (Example IA - Table 1).

Human gingival fibroblasts are used as normal cells control (Example 13 - Table 2). The cells are grown in Dulbecco's modified Eagles' medium (MEM) . supplemented with 10% foetal bovine serum, IX MEM vitamins, lXL-glutamine, IX ΝΞΑΑ3 (essential amino acids), IX gentamycin. They are incubated at 37JC in a humidified CO2 atmosphere. Approximately 1 to 3 x 105 cells are put into each well of a 24 well . microtiter plate containing 2 ml of the medium. PDP (potassium salt) in varying concentrations, is added.

After incubation, the cell viability is determined by removing aliquots from the wells over the time specified in Table 1. The viability is assessed . by the trypan blue exclusion test. Fresh medium is added in each well, each day to maintain the necessary growth conditions. The inhibition was calculated by comparing % of cells alive in phosphate buffer saline (PBS) vs. PDP. The data are summarized . in Table 1. - 10 TABLE 1 EFFECTS OF PDP ON MORINE MYELOMA (SPg LINE) CELLS NUMBERS OF CELLS x 105 % VIABLE TREATMENT N (AT 72 HOURS) CELLS Control (PBS) 4 8.93 + 0.14 100% PDP pH 7.0 100 mcg/ml 4 1.86 + 0.14 47 500 mcg/ml 4 1.33 + 0.03 33 1000 mcg/ml 4 1.07 + 0.17 29 2500 mcg/ml 4 0.48 + 0.15 12 These results show that compared to the buffer control, potassium salt of PDP is hignly cytotoxic and inhibitative to tbe murine myeloma (cancer) cells. Table 2 describes tbe effects on normal cells (human gingival fibroblast).

TABLE 2 EFFECTS ON PDP ON HUMAN GINGIVAL FIBROBLAST NUMBER OF VIABLE CELLS % VIABLE . TREATMENT N X 105 (AT 72 hours ) CELLS Control (PBS) 4 2.67 + 0.17 100 PDP pH 7.0 100 mcg/ml 4 2.61 + 0.16 98 500 mcg/ml 4 2.58 + 0.13 97 25 . 1000 mcg/ml 4 2.12 + 0.15 79 2500 mcg/ml 4 1.97 + 0.11 74 The data in Table 2 suggest no significant effect on cell growth at 100-500 mcg/ml of PDP but 30. that even on normal cells i, viability is reduced at 1000 and 2500 mcg/ml. It is noteworthy that the effect on the myeloma tumour cells (Table 1) even at high concentrations is more pronounced than the effect with the normal cells (Table 2).

. Similar results are obtained with lithium, sodium, magnesium, calcium, strontium, zinc and stannous salts of PDF, organic peroxydiphosphate as well as alkyl, adenylyl, guanylyl, cytosylyl, thymylyl esters and tetramethyl ammonium salt of PDP.

. EXAMPLES 2A, 23 and 2C: The Effects of PDP, Potassium Pyrophosphate (KPP) and PBS (Phosphate Buffered Saline) on Tumour Development in Vivo Seventy-five genetically identical Balb/C mice having an average weight of 20 grams + 3 grams groups . of 25 animals each: (a) control treated with phosphate buffer saline (PBS); (b) treated with potassium peroxydiphosphate (PDP) and PBS, pH 7.0; and (c) potassium pyrophosphate (KPP) and PBS as a phosphate control. Each animal receives 0.2 ml of . pristane intraperitoneally (I.P.) to prime animals for malignant SPg cells (murine myeloma carcinoma tumour cells) implantation. After three weeks, the animals are put on oral ingestion treatment regime as follows: Group (a) receiving via I.P. 0.2 ml of PBS, . group (b) receiving 2.0 mg PDP suspended in 0.2 ml of PBS, and group (c) receiving 2.0 mg, of KPP in 0.2 ml of PBS, for three consecutive days. Forty-eight hours after the third injection, each animal is inoculated (I.P.) with-2 to 3 x 106 cells of SP2 3θ- (mice tumour cells, murine myeloma). Thereafter, the animals are given their respective materials, once daily for 5 days/week. That is, (a) PBS, (b) PDP or (c) KPP. The animals are scored for tumour development and death each week. The data is . analyzed using the Hantel-Haenszel procedure (Statistical Aspects of the Analysis of Data from Retrospective Studies of Disease, J. National Cancer Institute, Vol. 3, 719-748, 1959). The data in Tables 3, 4 and 5 indicate that PDP is significantly . effective in controlling tumour development in mice when compared to P3S or KPP, thereby evidencing that the effects in inhibiting tumour development is due to the provision of active oxygen species and not phosphate .

. . . TABLE 3 PBS* VS. KPP** TEN ί WEEK TUMOUR DEVELOPMENT STUDY NO TUMOUR AT RISK WEEK TREATMENT TUMOUR AND DEATH 5. 1-4 PBS 11 14 25 KPP 10 15 25 5 P3S 4 10 14 KPP 4 11 15 6 PBS 5 5 10 10. KPP 2 9 11 7 PBS 2 3 5 KPP 4 5 9 8 PBS 0 3 3 KPP 1 4 5 15. 9 PBS 2 1 3 KPP 3 1 4 10 PBS 1 0 1 KPP 0 1 1 20 . Mantel-Haenszel chi-square = 0.36 with 1, d.f. , P = 0.55 Odds ratio - 1. 34 These results are not significant and evidence no significant difference between PBS and KPP in reduce 25. tumour development in the animal.

*PBS = Phosphate buffer saline **KPP = Potassium pyrophosphate.

. TABLE 4 PBS* VS. PDP** TEN WEEK TUMOUR STUDY DEATH NO TUMOUR AT RISK WEEK TREATMENT TUMOUR AND 5 . 1-4 PBS 11 14 25 PDP 2 23 25 5 P3S 4 10 14 PDP .4 19 O T I* 6 PBS 5 5 10 10. PDP 5 14 19 7 PBS 2 2 5 PDP 2 13 14 8 PBS 0 3 3 PDP 2 10 12 15. 9 PBS 2 1 PDP 3 7 10 10 PBS 1 0 . l PDP 1 6 —Ί / 20. Mantel -Haenszel chi square = 10.40 with 1, d. f. , P - 0.001.

Odds ratio = 3.66.

These data indicate that the PBS control group develop tumours significantly sooner than PDP treated animals . (P = 0.001).

*PBS = Phosphate buffer saline **PDP = Potassium peroxydiphosphate.

. TABLE 5 KPP* vs. ρρρ*** TEN WEEK TUMOUR STUDY WEEK TREATMENT TUMOUR AND DEATH NO TUMOUR . AT RI31- 5. 1-4 KPP 10 15 25 PDP 2 23 25 5 KPP 11 15 PDP 4- 19 23 6 KPP 2 9 11 10 . PDP 5 14 19 7 KPP 4 5 9 PDP 2 14 14 8 KPP 1 4 5 PDP 2 10 12 15. 9 KPP 3 1 A —τ PDP 3 7 10 10 KPP 0 1 1 PDP 1 6 7 Mantel-Haenszel chi square = 5.86 with 1, d.f. , P = 20. 0.02 • Odds ratio = 2. 60. The data indicated that the KPP group develops tumours significantly sooner than PDP treated animal: (P - 0.001).

. * KPP = Potassium pyrophosphate *** PDP = Potassium peroxydiphosphate.

. Similar results can be observed when each of P3S, KPP and PDP are administered intramuscularly and intravenously in the same concentrations in PBS or orally in a concentration of 1 mg/ml (0.1%) in a . stable carrier of 45 parts of N-butyl stearate, 20 parts of carnauba wax, 25 parts of stearic acid and 10 parts of cellulose acetate phthalate.

Similar results are obtained with other inorganic salts of PDP, particularly lithium, sodium, . magnesium, calcium, strontium, zinc and stannous salts. Organic compounds of PDP, particularly C^_]_2 alkyl, adenylyl, guanylyl, cytosylyl, thymylyl esters and tetramethyl ammonium salts are also effective in countering growth of murine myeloma malignant tumour . cells.

EXAMPLE 3 500 parts of potassium peroxydiphosphate and 641 parts of mannitol are blended and wet with 32.5 parts of a 10% solution of sorbitol to form a wet . granulate, which is dried at 49°C and screened through a 12 mesh, O.S. Sieve (1.68 mm screen openings). 35 parts of magnesium stearate is then added as a binder and tabletted granules are formed by compressing the composition on a tablet . compressing machine.

The tablets are coated with an enteric coating solution of the following composition: Cellulose acetate phthalate 120 parts Carnauba wax 30 parts Stearic acid 10 parts 95% ethanol 450 parts Acetone Q.S. to 1000 parts The coating is carried out hy a pouring procedure in a conventional coating pan.

When the tablets thus formed are ingested, they pass through the stomach without breakdown and the . coating is then dissolved by intestinal fluids.

EXAMPLE 4 Deionized distilled water is stabilized at atmospheric pressure for 20 minutes in an autoclave. After cooling, it is tested for non-pyrogenicity . using the Limulus Amoebocyte Lysate (LAL) as described by Tsuji et al in Pharmaceutical Manufacturing, October, 1984, pages 35-41. 50 parts of potassium peroxydiphosphate, sodium chloride in amount corresponding to 0.9% of solution and 0.1 M . phosphate buffer containing KH2PO4 and NagHPOz., pH 9.4 are added to the pyrogen-free sterile water. The solution is then sterilized by passing it through a 0.5 micropore filter and is then packed in sterile files.

. Although the invention has been described with reference to specific examples, it will be apparent to one skilled in the art that various modifications can be made thereto which fall within its scope.

Claims

1. The use of peroxydiphosphoric acid, or a pharmaceutically acceptable compound derivative thereof, in the preparation of an anticancer agent.

2. A sterile preparation comprising peroxydiphosphoric acid, or a pharmaceutically acceptable compound derivative thereof, and a pharmaceutically acceptable carrier, which is buffered phosphate saline.

3. A tablet preparation comprising peroxydiphosphoric acid, or a pharmaceutically acceptable compound derivative thereof, coated with a material which resists breakdown by gastric acids and which is broken down by intestinal fluids at a pH of from 5.5 to 10.

4. A composition comprising a dosage amount of from 0.1 to 10% by weight of a non-toxic water-soluble pharmaceutically acceptable compound derivate of peroxydiphosphoric acid dissolved or dispersed in a pharmaceutical carrier which pharmaceutical carrier is a buffered phosphate saline solution having a pH of about 7.0 to 7.4 or a coated tablet material which resists breakdown by gastric acids while being broken down by intestinal fluid at a pH of about 5.5 to 10.

5. A composition as claimed in Claim 4 wherein said pharmaceutical carrier is said buffered phosphate saline solution.

6. A composition as claimed in Claim 4 wherein said pharmaceutical carrier is said coated tablet material which resists breakdown by gastric acids while being broken down by intestinal fluid at a pH of from 5.5 to 10.

7. A composition as claimed in Claim 3 or 6 wherein the coating of the tablet comprises from 40 to 50 parts by weight of a fatty acid ester, from 15 to 25 parts by weight of a wax, from 20 to 30 parts by weight of a fatty acid and from 5 to 15 parts by weight of a cellulose ester. 19.

8. A composition as claimed in Claim 7 wherein the fatty acid ester is N-butyl stearate, the wax is carnauba wax, the fatty acid is stearic acid and the celluslose ester is cellulose acetate phthalate.

9. The subject matter as claimed in any one of Claims 1 to 8, wherein 5 the ron-toxic water-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid is a C^ alkyl, adenylyl, guanylyl, cytosylyl or thymylyl ester or a quaternary ammonium salt.

10. The subject matter as claimed in Claim 9 wherein the compound derivative is a C^ ester peroxydiphosphoric acid. 10.HThe subject matter as claimed in Claim 9 wherein the compound derivative is an adenosyl, guanylyl, cytosyl or thymyl ester of peroxydiphosphoric acid.

11.

12. A pharmaceutical pack containing a preparation comprising peroxydiphosphoric acid or a non-toxic water-soluble pharmaceutically 15. acceptable compound derivative thereof dissolved or dispersed within a pharmaceutical carrier which is a coated tablet which resists breakdown by gastric acids while being broken down by intestinal fluid at a pH of about 5.5 to 10, there being associated with the pack instructions to administer the preparation to a warm blodded animal host by oral 20. ingestion in a regimen which provides about 0.1-6 g per kg body weight of the warm blooded animal per day. 12

13. A method of preparing an anticancer agent, the method comprising forming a tablet of peroxydiphosphoric acid, or a pharmaceutically acceptable compound derivative thereof, coated with a material which 25. resists breakdown by gastric acids and which is broken down by intestinal fluids at a pH of from 5.5 to 10. 13

14. A method as claimed in Claim 13 wherein the coating of the tablet comprises from 40 to 50 parts by weight of N-butyl stearate, from 15 to 25 parts by weight of carnauba wax, from 20 to 30 parts by weight of 39. stearic acid and from 5 to 15 parts by weight of cellulose acetate phthalate. 14.

15. A method as claimed in Claim 13 wherein the non-toxic, water-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid is an alkali metal, alkaline earth metal, zinc or tin salt. 15A method as claimed in Claim 13 wherein the non-toxic water-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid is a C^ alkyl, adenylyl, guanylyl, cytosylyl or thymylyl ester, or a quaternary ammonium salt. 15 17. A method as claimed in Claim 15 wherein the salt is potassium peroxydiphosphate.

16. 18. A method as claimed in Claim 15 wherein the compound derivative is a a ^yl ester of peroxydiphosphoric acid.

17. 19. A method as claimed in Claim 16 wherein the compound derivative is an adenyly, guanylyl, cytosylyl or thymylyl ester % of peroxydiphosphoric acid.

18. 20. A pharmaceutical pack containing a preparation comprising a non-toxic water-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid, or the acid itself, dissolved or dispersed in a pharmaceutical carrier which has a physiological pH of from 7.0 to 7.4, there being associated with the pack instructions to administer the preparation systemically to a warm blooded animal host in a regimen which provides about 0.1-2 g per kg body weight of a warm blooded animal per day.

19. 21. A method of preparing an anticancer agent, the method comprising preparing a sterile solution or dispersion of peroxydiphosphoric acid, or a pharmaceutically acceptable compound derivative thereof, in an injectable carrier.

20. 22. A method as claimed in Claim 21 wherein the non-toxic water-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid is an alkali metal, alkaline earth metal, zinc or tin salt.

21. 23. A method as climed in Claim 21 wherein the non-toxic water-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid is a Cj jg alkyl, adenylyl, guanylyl, cytosylyl or thymylyl ester or a quaternary ammonium salt.

22. 24. A method as claimed in Claim 22 wherein the salt is potassium peroxydiphosphate.

23. 25. A method as claimed in Claim 23 wherein the compound derivative is a Cj_jg alkyl salt of peroxydiphosphoric ester.

24. 26. A method as claimed in Claim 23 wherein the compound derivative is an adenylyl, guanylyl, cytosylyl or thymylyl ester of peroxydiphosphoric acid.

25. 27. A method as claimed in Claim 21 wherein the carrier is physiologically acceptable pyrogen-free buffered phosphate saline.

26. 28. A method of making tabletted granules having a coating thereon which are not broken down during passage in the stomach and which coating is dissolved by intestinal fluids having a pH of from 5.5 to 10, which method comprises blending a non-toxic water-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid, or the acid itself, with a polyhydroxy sugar solid and wetting the blend with a polyhydroxy sugar compound solution, screening to size, blending a binding agent therewith, compressing to form tabletted granules and coating the tabletted granules by spraying a film of coating solution which is not inactivated by gastric acids and which is dissolved by intestinal fluids having a pH of from 5.5 to 10.

27. 29. A method of forming a solution of a non-toxic water-soluble pharmaceutically acceptable compound derivative of peroxydiphosphoric acid, or of the acid itself, suitable for stomach administration which comprises sterilizing deionized distilled water to be non-pyrogenic and then adding thereto a phosphate buffer and peroxydiphosphoric acid or its compound derivative, and sodium chloride.

28. 30. A suppository comprising peroxydiphosphoric acid, or a pharmaceutically acceptable compound derivative thereof, and a rectally or vaginally acceptable carrier.

29. 31. A pharmaceutical formulation containing peroxydiphosphoric acid, or 5 a pharmaceutically acceptable compound derivative thereof, and a pharmaceutically acceptable carrier, the formulation being other than in the form of a dentifrice, a mouthwash, chewing gum, a lozenge or a sorbitol-containing composition.

30. 32. A pharmaceutical formulation containing peroxydiphosphoric acid, or 10 a pharmaceutically acceptable compound derivative thereof, and a pharmaceutically acceptable carrier, the formulation being substantially free of humectants, ethanol and saccharides. 31 33. An anticancer preparation substantially as herein described with reference to any one of the Examples. 15 32 34. The use of peroxydiphosphoric acid, or a pharmaceutically acceptable compound derivative thereof, in the preparation of an anticancer agent substantially as herein described with reference to any one of the Examples. as defined in Claim 13 or Claim 21, 33. 35. A method of preparing an anticancer agent/substantially as 34.hereinbefore described with reference to either of Examples 3 or 4. 35. An anticancer agent whenever prepared by a method as claimed in any of Claims 13 to 19, 21 to 27 or Claim 35.