CS271461B2 - Method of pellet granules production with coating - Google Patents

Abstract

The solution concerns the method of production of tablet granules with coating that do not melt when passing through stomach. The coating is melted by intestinal fluids of pH ranging from 5.5 to 10. Its principle consists in the fact that 30 to 50 weight parts of non-toxic, water-soluble pharmaceutically suitable compound, derived from peroxydiphosphoric acid, where the derivate is the salt of alkaline metal, alkaline earth metal, zinc and tin, C1-12 alkyl ester, adenyl ester, guanyl ester, guanylyl ester, cytosylyl ester, thymyl ester or quaternary ammonium salt, is mixed with 45 to 60 weight part of polyhydroxy saccharide solution, mainly sorbitol solution, it is sorted according to size and mixed with 20 to 35 weight parts of binding agent like magnesium stearate. It is then pressed into the form of granules and these granules by means of spraying are coated with film of coating solution containing 15 to 25 weight parts of carnauba wax, 20 to 30 weight parts of stearic acid and 5 to 15 weight parts of acetate phthalate of cellulose in 400 to 900 weight parts of anorganic solvent that resists the gastric acids and is dissolved by intestinal fluids that have a pH value ranging from 5.5 to 10.

Description

BACKGROUND OF THE INVENTION The present invention relates to a process for the manufacture of coated tablet granules that do not break during gastric transit, wherein the coating dissolves by intestinal fluids at pH

Such tablet granules are useful in inhibiting tumor development in terms of tumor cells in vitro and actual tumor development in vivo in warm-blooded animals.

Cancer disease It is the result of malignant tumors. Much medical research has been devoted to reducing and overcoming the risk of cancer. To date, no cancer cure has been found. However, much of the mechanism by which warm-blooded animals prevent cancer-induced damage has been recognized. The present invention builds on this knowledge to provide a substance that inhibits tumor development and a method of inhibiting this development.

Cells contained in mammalian body fluids include lymphocytes, monocytes, macrophages, and polymorphic nuclear cells. These cells act as a natural safety system against tumor development in lower mammals such as rodents to mammals such as humans. In recent years, it has been found that certain subpopulations of lymphocytes or lymphoid cells, referred to as natural killer or NK cells, destroy tumor cells and thus prevent cancer development. Many findings suggest that NK cells have cytolytic activity relationship к formation of substances with active nitrogen such as hydrogen peroxide / Н ₂ 0 ₂ / ^ne ^ ° oxy-containing radicals, for example hydroxy anion / ОН / and superoxide anion ^{/ 0} ₂ 'Л

NK cells and active oxygen phenomenon are described by Herberman et al. in Science, Vol. 214. 298, Aug. 5, 1982, pp. 569-572; Nathan et al., Journal of Immunology, Vol. 129, No. 5, November 1982, pp. 2164-2171; and Mavier et al., Journal of Immunology, Vol. 132, No. 4, April 1984, pp. 1980-1986,

There are many compounds that release active oxygen. However, this factor alone does not mean that such compounds could be introduced into the body to complement NK cell function or where tumor formation is insufficient to provide NK cell function and inhibit tumor development. Compounds that release active oxygen usually evolve it rapidly, and to exhibit efficacy against tumor development in warm-blooded animals, such as humans, a slower and more delayed release is required. Until the present invention, this has not been effectively achieved. When oxygen is released too quickly, both tumor and normal cells can be attacked.

U.S. Pat. No. 4,041,149 issued in August 1977 to the author and co-authors of the present invention describes a composition in various forms, including dental tablets, that inhibit the formation of oral malodour in which the active ingredient is a peroxydiphosphate. The peroxydiphosphate compound differs from most oxygen generating compounds in that it does not produce an initial hydrogen peroxide jet. Rather, it releases hydrogen peroxide slowly so that when comparing equivalent concentrations to hydrogen peroxide, the amount of oxygen released by the peroxydisulfate once is equal to one tenth of the amount of available oxygen released by the hydrogen peroxide. Moreover, only about 50% of the active oxygen is released within 20 hours at 25 ° C in the presence of alkaline phosphatase or acid phosphatase, which are present in the bodies of warm-blooded animals including mice, rats, humans, etc.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a process for the manufacture of coated tablet granules that do not break during gastric passage, and which coat dissolves by intestinal fluids at pH 5.5 to 10 comprising mixing 30 to 50 parts by weight of non-toxic water soluble, pharmaceutically acceptable peroxydiphosphoric acid derivatives, the derivative being an alkali metal, alkaline earth metal, zinc and tin salt, a C 1-4 alkyl ester, an adenyl ester, a guanylyl ester, a cytosylyl ester, a thymyl ester or a quaternary ammonium salt, with 45 to 60 parts by weight solids, in particular mannitol, and the mixture is moistened with 25 to 35 parts by weight of a polyhydroxy-sugar solution, in particular sorbitol solution, sized, mixed and 20 to 35 parts by weight of a binder such as magnesium stearate, pressed into a mold

CS 271461 82

2;

tablet granules and these granules are spray coated with a film coating solution containing 15 to 25 parts by weight of carnauba wax, 20 to 30 parts by weight of stearic acid and 5 to 15 parts by weight of cellulose acetate phthalate in 400 to 900 parts by weight of an inorganic solvent that resists gastric acids and dissolves with intestinal fluids having a pH of 5.5 to 10 ·

The advantage of the present invention is that tumor growth is inhibited in tumor cells in vitro and the development of a true malignant tumor in vivo in warm-blooded animals such as rodents to humans *

A further advantage of the present invention is that a method of inhibiting tumor formation by introducing a slow release agent of a living host is provided. A composition comprising a dosage amount of about 0.1 to 10% non-toxic, water-soluble, pharmaceutically acceptable peroxydiphosphoric derivative compound may be used.

The peroxydiphosphate compound (РОР) is in the form of a non-toxic pharmaceutically acceptable compound which lies outside the salt disclosed in the aforementioned U.S. Patent 4,041,149.

The compounds include alkali metal salts (e.g., lithium, sodium and potassium), alkaline earth metals (e.g., magnesium, calcium and strontium), zinc and tin, as well as organic C 1-12 ^and alkyl, adenylyl, cytosylyl and thymylyl esters with peroxydiphosphates and also quaternary ammonium and similar salts ·

Alkali metal salts, especially potassium salts, are preferred among inorganic salts. Potassium peroxodiphosphate is an odorless fine free-flowing white non-hygroscopic crystalline substance having a molecular weight of 346.35 and an active oxygen content of 4.6%. 51% at 0 ° to 61 ° C, but insoluble in conventional solvents such as acetonitrile, alcohols, ethers, ketones, dimethylformamide, dimethylsulfoxide and the like. In a 2% solution in water, it has a pH of about 9.6 and a saturated solution thereof. pH about 10.9 · 10% solution in water at 25 ° C showed no loss of active oxygen after 4 months · At 50 ° C, 10% solution showed a loss of active oxygen of 3% in 6 months ·

Of the organic esters, those that provide hydrophobic properties, such as those having C _11-12 alkyl radicals, and those that facilitate rapid uptake of the peroxydiphosphate residue by cells such as adenylyl, guanylyl, cytosylyl, are preferred. and thymylyl ·

Pharmaceutical carriers suitable for oral administration are coated tablets formed from a material that resists gastric acid breakage at gastric pH (about 1-3), since peroxide phosphate would be inactivated by these gastric acids · Rather, carriers with tableted granules of peroxydiphosphate salts dissolve by intestinal fluids that have a higher pH (about 5.5 to 10) and do not inactivate peroxydiphosphate leaving it by the action of phosphatase enzymes present in humans or other warm-blooded animals ·

The desired tablet coating solution is formed from fatty acid esters such as n-butyl stearate (typically about 40 to 50, preferably about 45 parts by weight), a wax such as carnauba wax (typically about 15 to 25, preferably about 20 parts by weight) fatty acid, such as stearic acid (typically about 20 to 30 parts, preferably 25 parts by weight) and cellulose esters such as cellulose acetate phthalate (typically 5 to 15 parts, preferably about 10 parts by weight) and an organic solvent (typically about 400 to 900 parts) Other coating materials include shellac and copolymers of maleic anhydride and ethylene compounds such as polyvinyl methyl ether Such coatings are different from tablets that are broken in the oral cavity, in which the tablet material typically contains sel 80 to 90 parts by weight of mannitol and about 30 to 40% by weight parts of magnesium stearate ·

Tablets of peroxydiphosphate salt granules are formed by mixing about 30 to 50 parts by weight of a peroxydiphosphate salt with about 45 to 65 parts by weight of a polyhydroxy carbohydrate such as mannitol and wetting about 20 to 35 parts by weight of a solution of a polyhydroxy carbohydrate compound such as sorbitol. 20 to 35 parts by weight of a binder such as magnesium stearate and molding the granules into tablets on a tablet press · Tablet granules are coated by spraying a foam of the coating solution onto the tablets and dried to remove the solvent · Such tablets differ from dental tablets which are compressed from granules without special protective coating ·

The effective dose for peroxydiphosphate administration according to the prescribed regimen when administered orally is about 0.1 to 6 per kg of body weight per day when administered systemically, such as by intramuscular, intraperitoneal or intravenous injection, then the dose is about 0.1 to 2. g per kilogram of body weight per day ·

Physiologically acceptable non-pyrogenic solvents are suitable carriers for use in known methods for systemic administration · Phosphate buffered saline at a physiological pH of 7 to 7.4 is a preferred carrier for systemic administration · These solvents are different from the water-moistening carriers commonly used in dental compositions · Such a solution is usually prepared by sterilizing deionized distilled water, a control to ensure pyrogenicity using a lysate assay using Limulue amebocyte, described by Tsuji et al., Pharmaceutical Manufacturing, October 1984, pp. 35-41, and then adding a phosphate buffer (pH e.g. 8.5-10), produced in non-pyrogenic sterile water and about 1-100 mg of the peroxydiphosphate derivative and sodium chloride at a concentration of about 0.5-1.5% by weight ·

The solution may be filled into ampoules for use after it has been resterilized by passing through a microporous filter. Alternatively, other solutions such as Ringer's solution containing 0.66% sodium chloride, 0.03% potassium chloride and 0.033% chloride may be used. calcium ·

The peroxydiphosphate compound POP releases hydrogen peroxide slowly in the presence of phosphatase enzymes according to the following equation:

^{0 0 0} и n

И ¹¹ - foafatasv ¹¹² x -OPOOPO —rosraras ^ -------- OOPO>

⁴ I II

------------------ 0o H ₂ 0 ₂ + P0 ^_4-3 wherein X is a non-toxic pharmaceutically acceptable cation, or an organic ester residue forms.

Phosphatase for peroxydiphosphate disruption is present in saliva as well as in plasma, intestinal fluids and white blood cells. · Slow oxygen release It is particularly effective in replenishing the efficacy of NK cells against malignant tumor cells, corresponding to peroxydiphosphate therapy. When warm-blooded animals are treated with the PDP of the present invention, it is desirable to provide a regimen in which treatment is continued at least until tumors have subsided.

The following comparative examples illustrate the invention. · All amounts are by weight unless otherwise indicated.

Example 1

In vitro PDP tumor cytotoxicity study

This study investigates the effects of POP at various concentrations, including growth, Murine myeloma (SP ₂ lines) cells (Table 1). · Human gingival fibroplasts are used

As a normal cell control (Table 2). Cells are grown in Dulbecco's modified

Eagles medium supplemented with 10% fetal bovine serum, lx MEM vitamins, LXL-glutamine, 1 _X NEAA ₃ 1X gentamycin. Jen incubated at 37 ° C in a humidified C0 ₂ atmosphere.

02

Approximately 1 to 3 x 10 6 cells are placed in each well of a 24 well microtiter plate / containing 2 ml of medium. POP (potassium salt) is added at various concentrations.

After incubation, cell viability is determined by removing aliquots from the wells for the time specified in Table 1. The viability is determined by trypan blue exclusion assay. Fresh medium is added to each well each day to maintain the necessary growth conditions. Inhibition was calculated by comparing the percentage of living cells in phosphate saline (PBS) versus PDP. Data are summarized in Table 1

Table 1 effects of PDP on Marina myeloma cells (line SP 2)

Treatment N cell number x 10 ®% viable 72 hours / cells

control / PBS / 4 8.98 2 0.14 100 ALIGN! PDP pH 7/0 100 mcg / ml 4 1,86 * 0.14 47 500 mcg / ml 4 1,33 * 1,07 * 0.03 33 1000 mcg / ml 4 1.17 29 2500 mcg / ml 4 0,48 2 0.15 12

These results indicate that PDP potassium is highly cytotoxic and inhibitory to Murine myeloma (cancer cells) compared to oral control.

Table 2 shows the effects on normal cells (human gingival fibroplast).

Table 2

Effects of PDP on human gingival fibroplast

Treatment N number of viable X10 ^S cells / in 72 hours / % viable cells control / PBS / 4 2.67 - 0.17 100 ALIGN! PDP pH 7/0 100 mcg / ml 4 2.61 - 0.16 98 500 mcg / ml 4 2.58 ± 0.13 97 1000 mcg / ml 4 2.12 i 0.15 79 2500 mcg / ml 4 1.97 or 0.11 74 Data in the table 2 do not no significant effect on cell growth at 100 to 500 mcg / ml PDP, but u normal cell life decreases e at 1000 and 2500 mcg / ml.

Remarkably, the effect on myeloma tumor cells (Table 1) even at high concentrations is more prominent than the effect on normal cells (Table 2).

Similar effects are obtained with lithium, sodium / magnesium, calcium / strontium, zinc and ointment salts of PDP, organic peroxydiphosphates, as well as C1 alkyl, adenylyl, guanylyl, cytosylyl, thymylyl esters and tetramethylammonium salts of PDP.

Example 2

Effects of PDP, potassium pyrophosphate (КРР) and PBS (phosphate buffered saline) on in vivo tumor development of genetically identical Balb / C mice having an average weight of 20 g and 3 g in groups of 25 animals each:

a) control treated with phosphate buffered saline (PBS),

(b) potassium peroxydiphosphate (РОР) and PBS treatment group, pH 7.0;

c) potassium pyrophosphate treatment group KPP and PBS as phosphate control ·

Each animal receives 0.2 ml of intraperitoneal (1R) landing to prepare animals for the implantation of malignant SP ₂ cells (Murine myeloma carcinoma tumor cells). After 3 weeks, animals are placed on the oral treatment regimen as follows: Group A receives 0.2 ml PBS intraperitoneally, Group V receives 2.0 mg POP suspended in 2 ml PBS and Group C receives 2.0 mg KPP in 0 2 ml PBS for 3 consecutive days · hours after the third injection, each animal is inoculated (1Р / 2 to 3.10 with ⁶ SP 2 cells / murine myeloma tumor cells) · animals then receive the respective substances once a day for 5 days a week ·

(a) PBS,

(b) POP or

c) KPP.

Animals are monitored for tumor development and death every week. Data are analyzed using the Mantel-Haensze 1 procedure / statistical aspects of data analysis from retrospective disease studies, Journal National Cancer Institute, Volume 3, 719-748, 1959).

The data in Tables 3,4 and 5 show that PDP is significantly effective in suppressing tumor development in mice as compared to PBS or KPP, thus establishing that the effects of inhibiting tumor development are due to the effects of active oxygen-containing substances and not phosphates.

Table 3

PBS ^X vs. KPP ^XX

Ten-week study of tumor development

week treatment tumor and death no tumor in danger 1-4 PBS 11 14 25 KPP 10 15 Dec 25 5 PBS 4 10 14 KPP 4 11 15 Dec 6 PBS 5 5 10 2 9 11 7 PBS 2 3 5 KPP 4 5 9 8 PBS 0 3 3 KPP 1 4 5 9 PBS 2 1 3 KPP 3 1 4 10 PBS 1 0 1 KPP 0 1 1 Mantel-Haenszel Chi-Square ^B 0.36 s 1 - i d.f · P * = 0.55, ratio inequality = 0.36.

These results are not significant and there was no significant difference between PBS and KPP in reducing tumor development in animals.

x PBS «Phosphoric Acid AOL Solution xx KPP · Potassium pyrophosphate

Table 4 Ten-week trial of tumors PBS ^X vs. POP ** week tumor treated and no tumor at risk

1-4 PBS 11 14 25 9 PDP 2 23 25 5 PBS 4 10 14 - POP 4 19 Dec 23 6 PBS 5 5 10 PDP 5 14 19 Dec 7 PBS 2 2 5 PDP 2 13 14 !> 8 PBS 0 3 3 PDP 2 10 12 9 PBS 2 1 3 PDP 3 7 10 10 PBS 1 0 1 PDP 1 6 7

Mantel-Heanazel chi-square * 10.40 a 1, d * f «, Ρ · 0.001, | nerovnoati ratio «3.66

These data show that the PBS control group develops tumors significantly earlier than PDP-treated animals * 1 / P-0,001./ | x PBS · Phosphoric Acid Solution xx POP · sodium peroxydiphosphate.

7.

| Table 5

Oeaet weeks study of tumors

KPP * counter PDP *** week tumor treatment and death no tumor in danger

1-4 KPP 10 15 Dec 25 PDP 2 23 25 5 KPP 4 11 15 Dec PDP 4 19 Dec 23 β KPP 2 9 11 PDP 5 14 19 Dec 7 KPP 4 5 9 PDP 2 14 14

continuation of Table 5

week treatment tumor and death no tumor v nel 8 KPP 1 4 5 PDP 2 10 12 9 KPP 3 1 4 PDP 3 7 10 10 KPP 0 1 1 PDP 1 6 7

safety

Mantel-Haenszel chi-square and 5.86 with 1, df, P ^and o, 02.

Inequality ratio «2.60.

The data show that the KPP group develops tumors significantly earlier than the PDP treated animals (Р = 0.001).

x KPP »potassium pyrophosphate xxx PDP = potassium peroxydiphosphate.

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

Similar results are obtained with other inorganic salts of PDP, in particular lithium, sodium, magnesium, calcium, strontium, zinc and stannous salts. Organic compounds of PDP, especially C1-4 ₂ alkyl, adenylyl, guanylyl, cytosylyl, thymylyl esters and tetramethylammonium soles are also effective in reversing the growth of murine myeloma malignant tumor cells.

Example 3

500 parts of potassium peroxydiphosphate and 641 parts of mannitol are mixed and moistened with 32.5 parts of a 10% sorbitol solution to form a wet granulate which is dried at 49 ° C and sieved through a 1.68 mm (12 mesh) screen. . 35 parts of magnesium stearate are then added as a binder and tableted granules are formed by compression of the mixture on a tabletting machine.

The tablets were 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. up to 1000 parts

The coating is carried out by a casting process in a conventional coating pan.

When the thus formed tablets are swallowed, they pass through the stomach without disturbance and coating and then dissolve the intestinal fluids.

Example 4

The deionized distilled water is stabilized at atmospheric pressure for 20 minutes in an autoclave. After cooling, it is assayed for pyrogenicity using Limulus amebocyte (LAL) lysate as described by Tsuji et al. in Pharmaceutical Manufacturing, October

1984, pp. 35-41. Then 50 parts of sodium peroxydiphosphate, sodium chloride corresponding to 0.9% solution and 0.1 M phosphate buffer containing

KH ₂ PO ₄ and Na 2 HPO 4, pH 9.4, into pyrogen-free sterile water. The solution is then sterilized by passing it through a 0.5 micropore filter and then filled into sterile containers.

Although the invention has been described with reference to specific embodiments, it will be apparent to those skilled in the art that various modifications may be made within the scope of the invention.

Claims (1)

A process for the manufacture of coated tablet granules which do not break during gastric passage and which is dissolved by intestinal fluids at a pH of 5.5 to 10, characterized by mixing 30 to 50 parts by weight of a non-toxic, water-soluble, pharmaceutically acceptable compound, derived from peroxydiphosphoric acid, the derivative being an alkali metal, alkaline earth metal, zinc and tin salt, alkyl ester, adenyl ester, guanylyl ester, cytosylyl ester, thymyl ester or quaternary ammonium salt, 8 45-60 parts by weight of a polyhydroxy sugar solid, especially mannitol; moisturizes 25 to 35 parts by weight of a polyhydroxy-saccharide solution, in particular sorbitol solution, sized, mixed with 20 to 35 parts by weight of a binder such as magnesium stearate, pressed into tablet granules and spray coated with a 15 to 25% film coating solution 20 to 30 parts by weight of stearic acid and 5 to 15 parts by weight of cellulose acetate phthalate in 400 to 900 parts by weight of an inorganic solvent which resists gastric acids and dissolves by intestinal fluids having a pH of 5.5 to 10.