HK1015686B - Use of phenol substituted diphosphonates as antineoplastic agents - Google Patents

Description

Use of phenol-substituted bisphosphates as antitumor agents

The present invention relates to antitumor agents, in particular to the treatment of neoplastic diseases with phenol substituted gem-diphosphate derivatives. More specifically, the present invention provides the use of certain phenol-substituted gem-diphosphate derivatives for the preparation of pharmaceutical compositions for the treatment and prevention of cancer and metastasis, and in particular for the treatment and prevention of Ras oncogene-dependent cancer and metastatic invasion.

Most of the existing anticancer drugs are cytotoxic compounds which lack specificity for killing tumor cells and thus affect normal cells, thereby generating toxic and side effects. There is therefore a need for the development of more specific agents for cell signaling pathways that result in inhibition of cancer cell proliferation without affecting normal cell proliferation (oncolytic drugs, J.R.Prous, Annual letters of specialty drugs, 1994 edition, page 459 and methods for using Ras oncogenes in cancer chemotherapy, G.Bolton et al, Annual Reports in medical Chemistry, 1994: 29: 165-17).

Mutations in the Ras tumor gene have been shown to be present in a variety of human tumors and these mutations can contribute to 1/5 in all human cancers. It has been found that more than 50% of colon cancers and 90% of pancreatic cancers are caused by it. Ras mutations are therefore thought to play an important role in triggering the development and progression of cancer (J.L.Bos, cancer research 1989; 49: 4682-. It has also been confirmed now that mutant forms of Ras protein are only present in tumors and not in normal tissues of cancer patients. Therefore, attractive therapeutic agents are those that block the activity of Ras mutations that convert normal cells into cancer cells and further promote the development of cancer cells and tumors.

U.S. Pat. No. 5,043,330(1991), corresponding to European patent No. 0,339,237, discloses a class of phenol-substituted gem-diphosphate derivatives and their use as lipid-lowering agents, such as for the treatment of cardiovascular diseases.

The applicant has now surprisingly found that such bisphosphates disclosed in US 5,043,330 are capable of specifically inhibiting the proliferation of cancer cells and inducing apoptosis of cancer cells without being toxic to normal cells. Thus, in one aspect, the present invention provides the use of a compound for the manufacture of a medicament for the treatment of a neoplastic disease, said compound having the following general formula (I):wherein: -Z¹、Z²、Z³And Z⁴Are identical OR different and are-OR, where R is H, straight-chain, branched OR alkyl having from 1 to 8 carbon atoms, -OM, where M is a cation, -NR₂Wherein R has the same meaning as defined above, -Z¹、Z²And Z³、Z⁴May form an alkylene dioxycycle having from 2 to 8 carbon atoms, -X¹、X²Are identical or different and are H, a halogen atom, a linear, branched or cyclic alkyl or alkoxy radical having 1 to 8 carbon atoms, X³Is H, an alkyl radical R having from 1 to 4 carbon atoms¹Acyl C (O) R¹Carbamoyl C (O) NHR¹Wherein R is¹As described above, X³O and two other substituents X¹Or X²2-may form an alkylenedioxycycle containing from 1 to 4 carbon atoms, -a is-CH ═ CH-CH₂-、-(CH₂)_n-、-O(CH₂)_n-、-S-、-SO₂-、-S(CH₂)_n-、-SO₂(CH₂)_n-, where n is an integer from 1 to 7, or A and B form a compound of the formula- (CH ═ CH)_k-(CH₂)_d-CH ═ alkylene, where k is 0 or 1 and d is an integer from 0 to 4, -B is H, an alkyl group containing 1 to 4 carbon atoms, -t is 0 or 1, with the proviso that when a is- (CH ═ CH)_k-(CH₂)_d-CH ═ where k and d are as described above, when t is 0.

The compounds of formula (I) may be present as salts, and the compounds of formula (I) below include salt forms of the compounds, unless otherwise indicated. Examples of salts are compounds of the general formula (I) in which Z¹、Z²、Z³And Z⁴Is composed of OM or an amino onium NR₄Wherein M is an alkali metal or alkaline earth metal ion or R has the same meaning as defined above.

In another aspect, the invention provides the use of a compound of formula (I) as hereinbefore defined for the preparation of a medicament for the treatment of solid tumours, for example colon, pancreas, thyroid, lung, breast, brain and neck tumours.

In another aspect, the invention provides the use of a compound of formula (I) as defined above for the preparation of a medicament for the treatment of tumours of the haematopoietic and immune system such as lymphomas and leukaemias.

In a further aspect, the present invention provides the use of a compound of formula (I) as hereinbefore defined for the manufacture of a medicament for the treatment of patients with primary tumour metastases.

In a further aspect, the invention provides the use of a compound of formula (I) as hereinbefore defined in the manufacture of a medicament for preventing transformation of normal cells or for inhibiting metastatic invasion of cancer cells to normal cells.

In a further aspect, the invention provides a method of treating a neoplastic disease or preventing cancer metastasis, particularly Ras-dependent cancer, which comprises administering to a patient suffering from or potentially developing cancer a therapeutically effective amount of a compound of formula (I) as defined above.

Also within the scope of the invention is a method of selectively removing cancer cells, the method comprising treating a mixture of cancer cells and normal cells from a patient in vitro with a compound of formula (I), and then transferring the cells to the patient. Thus, according to the method, blood, plasma or other body fluid can be withdrawn from a patient, treated ex vivo with a compound of formula (I) and then transferred to the patient.

In the compounds of the general formula (I),Z¹、Z²、Z³and Z⁴Examples of the group include hydroxy, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy and tert-butoxy. The preferred group is isopropoxy.

Preference is given here to Z¹、Z²、Z³And Z⁴The radicals being identical, a particularly preferred embodiment of the present invention is Z¹、Z²、Z³And Z⁴The groups are all isopropoxy.

Group X¹And X²Examples of (b) include hydrogen, straight or branched chain alkyl groups and alkoxy groups having 1 to 5 carbon atoms, with 1 to 4 carbon atoms being particularly preferred. X¹And X²Preferred groups of (A) are methyl, ethyl, n-propyl, isopropyl, sec-butyl, tert-butyl, methoxy and ethoxy, tert-butyl being particularly preferred.

Group X³Examples of (2) include hydrogen, C_1-4Alkyl and C_1-4Alkanoyl, with hydrogen being particularly preferred herein.

The compounds of the general formula (I) include phenol-substituted alkylene diphosphates (Ia) and phenol-substituted alkenylene diphosphates (Ib).Wherein X¹、X²、X³、A、B、k、d、Z¹、Z²、Z³And Z⁴As described above.

Compounds of structure (Ia) include, for example, those wherein: -X¹And X²Are identical or different and are alkyl having 1 to 8 carbon atoms, -X³Is hydrogen, -A is CH ═ CH-CH₂(CH₂)_n，S，SO₂，S-(CH₂)_n，SO₂-(CH₂)_nWherein n is 1-7, -B is hydrogen or C₁-C₄Alkyl, -Z¹、Z²、Z³And Z⁴Are identical or different and are OH, alkoxy having 1 to 8 carbon atoms or Z¹，Z²And Z³，Z⁴One or both of the pairs is an alkylenedioxy group having from 2 to 8 carbon atoms.

Compounds of structure (Ib) include, for example, those wherein: -X¹And X²Are identical or different and are alkyl having 1 to 8 carbon atoms, -X³Is hydrogen, -k is 0 or 1, d is 0 to 4, -Z¹、Z²、Z³And Z⁴Are identical or different and are OH, alkoxy having 1 to 8 carbon atoms or Z¹、Z²And Z³、Z⁴One or both of the pairs is an alkylenedioxy group having from 2 to 8 carbon atoms.

Specific examples of compounds of formula (I) useful in the present invention include the compounds in tables 1a and 1 b.

The present invention provides novel uses of gem-bisphosphonates of formula (I) for the treatment of neoplastic diseases and for the prevention of cancer and metastasis, in particular those Ras-dependent cancers. In a particularly preferred embodiment, it provides a novel use of compound 1 of formula (I) wherein X is¹And X²Are both tert-butyl radicals, in the 3-and 5-positions, respectively, X³Is H, in the 4-position, A is CH₂B is H, t is 1, Z¹、Z²、Z³、Z⁴Are all isopropoxy. The compound 1 has the following structure, general formula and physical and chemical properties:2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -ethylene-1, 1-bisphosphate tetraisopropyl ester, C₂₈H₅₂O₇P₇，mp＝104-105℃。

The compounds of the invention can be prepared according to the procedures described in EP 0339237A, which is incorporated herein by reference in EP 0339237A.

Some of the analogs are novel. Thus, in another aspect, the present invention provides novel compounds selected from: 2- (3, 5-diisopropyl-4-hydroxyphenyl) -ethenylene-1, 1-tetraisopropyl diphosphate 2- (3, 5-diisopropyl-4-hydroxyphenyl) -ethylidene-1, 1-tetraisopropyl diphosphate 2- (3, 4, 5-trimethoxyphenyl) -ethenylene-1, 1-tetraisopropyl diphosphate 2- (3, 4, 5-trimethoxyphenyl) -ethylidene-1, 1-tetraisopropyl diphosphate 2- (3-tert-butyl-4-hydroxy-5-methylphenyl) -ethenylene-1, 1-tetraisopropyl diphosphate 2- (3-tert-butyl-4-hydroxy-5-methylphenyl) -ethylidene-1, tetraisopropyl 1-diphosphate 2- (3-ethoxy-4-hydroxyphenyl) -vinylene-1, 1-tetraisopropyl 2- (3-ethoxy-4-hydroxyphenyl) -ethylene-1, 1-diphosphate 2- (3, 5-di-tert-butyl-4-methoxyphenyl) -vinylene-1, 1-diphosphonate tetraethyl 1- (3, 5-di-tert-butyl-4-hydroxyphenyl) -butylene-2, 2-diphosphonate tetraisopropyl

The compounds of formula (I) may be administered orally or through other mucosal surfaces (e.g., nasal, buccal, bronchial or rectal mucosa), transdermally, or by injection (e.g., subcutaneous, intraperitoneal, intravenous or intramuscular injection).

When the compounds of formula (I) are intended for oral administration, they may be formulated, for example, as tablets, capsules, granules, pills, dragees, lozenges, powders, solutions, emulsions, syrups, suspensions or any other preparation suitable for oral administration. If desired, the oral dosage form may be coated with one or more sustained release coatings to control the release of the active compound at specific locations in the intestinal tract.

The compounds of formula (I) and pharmaceutically acceptable solubilizers, diluents or carriers can be formulated in standard manner into tablets and other solid or liquid dosage forms. Examples of solubilizers, diluents or carriers include sugars such as lactose, starch, cellulose and its derivatives, powdered tragacanth (tracaganth), malt, gelatin, talc, stearic acid, magnesium stearate, calcium sulfate, vegetable oils, polyols such as glycerol, propylene glycol and polyethylene glycol, alginic acid and alginates, agar, pyrogen-free water, isotonic saline, phosphate buffered solutions and optionally other pharmaceutical excipients such as disintegrants, lubricants, wetting agents such as sodium lauryl sulfate, colorants, flavors and preservatives, and the like.

Capsules can be of the hard or soft type, containing the active compound in solid, liquid or semi-solid form. Typically such capsules are formed from gelatin or equivalent and may or may not be coated. If it is desired that the capsule releases the active ingredient upon entry into the intestine through the stomach, the capsule may be provided with a pH sensitive coating to accommodate dissolution in the pH of the duodenum or ileum. Examples of such coatings include Eudragits, the use of which is well known in the art.

Injections typically consist of a suitable solubilizing agent, such as a detergent, which also includes compounds and excipients such as buffers which provide an isotonic solution of the correct physiological pH. Injection solutions are generally pyrogen-free and provide a sealed vial or ampoule containing a unit dose of the compound.

A unit dose of a compound of the invention will generally contain from 0.1% to 99% by weight of active material, more typically from 5% to 75% active material. By way of example, a unit dose may contain from 1mg to 1g of the compound, more typically from 10mg to 500mg, for example from 50mg to 400mg, typically a dose of between 100mg and 200 mg.

The compounds of the present invention will be administered in an amount effective to provide the desired therapeutic effect. The concentration which provides the desired therapeutic effect will vary depending on such factors as the precise nature of the disease, the size, weight and age of the patient and the severity of the disease.

Although in certain cases the severity of the disease being treated will require administration of an amount of the compound that causes some symptoms of toxicity, it is preferred that the dose administered is non-toxic to the patient.

In general, the compounds of the invention are administered in an amount of from 0.01mg/kg to 100mg/kg body weight, more preferably in an amount of from 0.1mg/kg to 10mg/kg body weight and especially from 1mg/kg to 5mg/kg body weight. For a human with an average body weight of 70kg, the daily dosage of the compound of the present invention is generally from 70mg to 700 mg. Such doses are administered, for example, two to four times per day. However, the size of the dose and the frequency of administration are at the discretion of the physician treating the patient.

Example K is intended to detail a representative formulation for the inventors' preparation of compound 1 capsules.

The pharmacological activity of the compound can be confirmed by adopting human bladder cancer T24(H-Ras) oncogene to transfect NIH 3T3 cell clone and an in vitro screening model. This cell line (PAP2) has been screened for Ras-dependent activity based on its high level of Ras expression and associated high level of metastatic capacity (S.A. Hill et al, J.Korea cancer institute 1988, 80: 484-490 and A.Champers et al, 1990 in invasion and metastasis; 10: 225-240). The PAP2 cell line has been shown to increase Ras-dependent expression of cathepsins, cysteine proteases involved in the metastatic process (A. Chambers et al, oncogene mology, 1992; 5: 238-245). Thus, PAP2 cells were implicated in the pathological process of human cancer. These cells were used in vitro assays to observe the effect of compounds on cell proliferation, proteolytic activity (metastasis) and flagellin (apoptosis). When these cells were injected subcutaneously (s.c.) into immunodeficient (nude) mice, solid tumors rapidly formed and then the anticancer activity of the test compounds was measured in vivo after oral administration of the test compounds.

The results of a series of in vitro and in vivo tests led to the discovery of the present invention that representative compounds of formula (I) and in particular Compound 1,

-inhibiting the growth of cancer cells in tissue culture,

-inducing apoptosis of cancer cells in tissue culture,

inhibition of proteolytic enzymes in cancer cells involved in metastasis and

shows anticancer activity in nude mice with solid tumors.

The test results shown in tables 1-7b provide evidence that compounds of general formula (I), in particular compound 1, are potentially useful in the treatment of neoplastic diseases including cancers of the hematopoietic and immune systems such as lymphoid and leukemias, and cancers of the pancreas, colon, breast, thyroid, brain, lung, head and neck. The recently discovered anti-cancer activity of the compounds of formula (I) is unexpected and independent of their lipid-lowering activity previously reported.

The results are expressed as averages. Significant differences were assessed using the Student's t-test of unpaired data.

Example 1 inhibition of Ras-dependent cell proliferation with Compounds of formula (I)

Screening a series of compounds of formula (I) to determine the maximum active compound and the relationship of structure to activity. Inhibition of proliferation of PAP2 cells was selected as the initial screening test.

Briefly, at 3 × 10 per hole⁴The concentration of PAP2 cells were seeded on a 24-well plate and allowed to attach for 24 hours. Test compounds were added at final concentrations of 10 and 20 μ M in 1% ethanol solution. After 48 hours of incubation, cells were trypsinized and visible cells (except trypan blue) were counted. The results obtained for the series of compounds (Ia) and (Ib) are shown in tables 1a and 1b, respectively.

The compounds to be screened in the assay can be synthesized according to the procedures described in U.S. Pat. No. 5,043,330(1991), corresponding to European patent 0339237. Some examples (examples a-J) are provided to illustrate in more detail the synthesis of the novel derivatives according to the procedures described in the prior art above.

TABLE 1A Effect of phenol-substituted gem-bisphosphonates (Ia) on PAP2 cells

Compound (I)	X1	X2	X3	A	B	Z1	Z2	Z3	Z4	Cells (% count control)
												10μM	20μM
										1	3-t-Bu			5-t-Bu	4-H	CH2	H	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr	-70	-100
2	3-t-Bu	5-t-Bu	4-H	CH2	H	OH	OH	OH	OH			-31	-45
										3	3-t-Bu			5-t-Bu	4-H	CH2	H	OMe	OMe	OMe	OMe	-21	-17
4	3-t-Bu	5-t-Bu	4-H	CH2	H	OEt	OEt	OEt	OEt			-30	-22
										5	3-t-Bu			5-t-Bu	4-H	CH2	H	On-Pr	On-Pr	On-Pr	On-Pr	-22	-89
6	3-t-Bu	5-t-Bu	4-H	CH2	H	On-Bu	On-Bu	On-Bu	On-Bu			-26	-51
										7	3-s-Bu			5-s-Bu	4-H	CH2	H	OEt	OEt	OEt	OEt	-6	+3
8	3-s-Bu	5-s-Bu	4-H	CH2	H	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-21	-45
										9	3-i-Pr			5-i-Pr	4-H	CH2	H	OEt	OEt	OEt	OEt	-16	-31
10	3-i-Pr	5-i-Pr	4-H	CH2	H	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-20	-51
										11	3-t-Bu			5-Me	4-H	CH2	H	OEt	OEt	OEt	OEt	-14	-37
12	3-t-Bu	5-Me	4-H	CH2	H	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-3	-27
										13	3-t-Bu			5-t-Bu	4-H	S	H	OEt	OEt	OEt	OEt	-19	-39
14	3-t-Bu	5-t-Bu	4-H	S	H	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-53	-90
										15	3-OMe			5-OMe	4-Me	CH2	H	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr	-23	-5
16	3-OEt	5-H	4-H	CH2	H	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-2	+5
										17	3-t-Bu			5-t-Bu	4-H	CH2	H	OEt	OEt	On-Bu	On-Bu	-6	-95
18	3-t-Bu	5-t-Bu	4-H	CH2	H	OEt	OEt	Oi-Pr	Oi-Pr			-46	-59
										19	3-t-Bu			5-t-Bu	4-H	CH2	Et	OEt	OEt	OEt	OEt	-21	-36
20	6-Cl	34-O	CH2	CH2	H	OEt	OEt	OEt	OEt			+20	-9
										21	3-t-Bu			5-t-Bu	4-H	CH2	H	O(CH2)3	O	O(CH2)3	O	-32	-28
22	3-OMe	5-OMe	4-H	CH2	H	OEt	OEt	OEt	OEt
										23	3-OMe			5-OMe	4-H	CH2	H	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr
24	3-t-Bu	5-t-Bu	4-H	CH2	Et	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-25	-40

TABLE 1b Effect of phenol-substituted gem-bisphosphonates (Ib) on PAP2 cells

Compound (I)	X1	X2	X3	k	d	Z1	Z2	Z3	Z4	Number of cells (% control)
												10μM	20μM
										25	3-t-Bu			5-t-Bu	4-H	0	0	OH	OH	OH	OH
26	3-t-Bu	5-t-Bu	4-H	0	0	OMe	OMe	OMe	OMe			-33	-17
										27	3-t-Bu			5-t-Bu	4-H	0	0	OEt	OEt	OEt	OEt	-48	-45
28	3-t-Bu	5-t-Bu	4-H	0	0	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-28	-46
										29	3-t-Bu			5-t-Bu	4-H	0	0	On-Pr	On-Pr	On-Pr	On-Pr	-51	-63
30	3-t-Bu	5-t-Bu	4-H	0	0	On-Bu	On-Bu	On-Bu	On-Bu			-13	-31
										31	3-s-Bu			5-s-Bu	4-H	0	0	OEt	OEt	OEt	OEt	-30	-44
32	3-s-Bu	5-s-Bu	4-H	0	0	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-13	-54
										33	3-i-Pr			5-i-Pr	4-H	0	0	OEt	OEt	OEt	OEt	-5	+13
34	3-i-Pr	5-i-Pr	4-H	0	0	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-26	-62
										35	3-t-Bu			5-Me	4-H	0	0	OEt	OEt	OEt	OEt	-14	-14
36	3-t-Bu	5-Me	4-H	0	0	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-24	-7
										37	3-OMe			5-OMe	4-H	0	0	OEt	OEt	OEt	OEt	-3	-28
38	3-OMe	5-OMe	4-H	0	0	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr
										39	3-OMe			5-OMe	4-Me	0	0	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr	-17	-34
40	3-OEt	5-H	4-H	0	0	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-30	-23
										41	3-t-Bu			5-t-Bu	4-H	1	0	OEt	OEt	OEt	OEt	-40	-77
42	3-t-Bu	5-t-Bu	4-H	1	0	Oi-Pr	Oi-Pr	Oi-Pr	Oi-Pr			-36	-71
										43	3-t-Bu			5-t-Bu	4-H	0	0	OEt	OEt	On-Bu	On-Bu	-18	-54
44	3-t-Bu	3-t-Bu	CH2	0	0	OEt	OEt	Oi-Pr	Oi-Pr			-17	-28
										45	3-t-Bu			5-t-Bu	4-Me	0	0	OEt	OEt	OEt	OEt	-10	-33
46	H	3，4-	OCH2	0	0	OEt	OEt	OEt	OEt			-22	-26
										47	H			3，4-	(OCH2)2	0	0	OEt	OEt	OEt	OEt	-11	-30

Example 2 in vitro test results inhibition of Ras-dependent cell proliferation cell culture

5% CO at 37 ℃ in Dulbecco's Modified Eagle Medium (DMEM)₂H-Ras-transfected NIH 3T3 cells (PAP2) (S.A. Hill et al; J. national cancer institute 1988; 80: 484-490) were grown in an atmosphere in which the medium contained 25mM HEPES and 10% fetal bovine serum. PAP2 cells were trypsinized and cultured twice more before fusion. 1. Inhibition of cell proliferation

The effect of compound 1 on cell proliferation was monitored by two methods:

cytometry with a hemocytometer and pooled DNA assays

-estimating the number of cells by colorimetry. 1.1 cell count and DNA content

Briefly, PAP2 cells were plated at 3 × 10 per well 4 hours prior to the addition of increasing concentrations of test compound⁴Is plated on a 24-well plate. The cells were trypsinized the next day later. Aliquots of the cell suspension were counted using a hemocytometer. The retained cells were dissolved in 0.01N NaOH and the DNA concentration was determined by fluorescence spectrophotometer using 4, 6-diamino-2-phenylindole as fluorescent dye and calf thymus DNA as standard.

TABLE 2a

Inhibition of PAP2 cell proliferation by compound 1

(cell number/well)

Concentration of Compound 1
								0	0.1μM	0.5μM	1.0μM	5.0μM	10μM
Cell number/well sem% change p	2737509437	2562507465-60.196	2087504270-240.001	18625015861-320.003	1025005951-630.001	712508260-740.001

The calculated IC50 value for inhibition of PAP2 cell growth by compound 1 was 1.02 μ M.

TABLE 2b

Reduction of Compound 1-produced DNA concentration in PAP2 cells in culture

DNA (mg/well)

Concentration of Compound 1
								0	0.1μM	0.5μM	1.0μM	5.0μM	10μM
DNA (mg/well) sem% change p	4.620.14	4.400.18-50.375	4.040.42-130.240	1.990.14-570.001	1.660.07-640.001	0.340.04-930.001

The calculated IC50 value for the reduction in PAP2 DNA content of Compound 1 was 2.77. mu.M.

The results in tables 2a and 2b show that the compounds of general formula (I), in particular compound 1, are able to inhibit the growth of PAP2 cells in culture. 1.2. Colorimetric MTT assay

Using a vaccine essentially as described in t.mosman in journal of immunization methods 1983; 65: the MTT assay described in 55-63 estimates cell number.

Briefly, 5 hours before addition of test drug, at 1 × 10 per well⁴The PAP2 cells were seeded onto 96-well dishes (Falcon). After 24 hours, 48 hours or 72 hours of incubation, 10. mu.l of MTT [3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide in PBS at 5mg/ml was added to each well]And incubated at 37 ℃ for 4 hours. The medium was then removed and 100. mu.10.04N HCl isopropanol solution was added to each well. The absorbance of the conversion dye at 570nm was determined using a microplate counter with the background at 620nm subtracted.

The calculated IC50 value of compound 1 for inhibition of proliferation of PAP2 cells was 8.05 μ M as determined by MTT method (after 72 hours incubation). The results in tables 3a, 3b and 3c obtained with another assay confirm that the compounds of general formula (I), in particular compound 1, are able to inhibit the growth of PAP2 cells in culture.

Effect of Compound 1 on PAP2 cell proliferation measured by MTT assay

TABLE 3a

Incubation of Compound 1 for 24 hours

(OD at 570 nm)

Concentration of Compound 1
								0	1.0μM	2.5μM	5μM	10μM	25μM
ODsem% change p	39.92.10	39.54.6-10.942	37.61.2-60.370	35.91.0-100.109	28.91.1-280.001	21.41.0-460.001

TABLE 3b

Compound 1 incubation for 48 hours

(OD at 570 nm)

Concentration of Compound 1
								0	1.0μM	2.5μM	5μM	10μM	25μM
Mean value sem% change p	71.35.20	66.54.9-70.516	59.42.5-170.059	52.83.1-260.009	37.31.9-480.001	15.10.7-790.001

TABLE 3c

Compound 1 incubation for 72 hours

(OD at 570 nm)

Concentration of Compound 1
								0	1.0μM	2.5μM	5μM	10μM	25μM
Mean value sem% change p	366.030.3	336.819.0-80.427	334.418.6-90.388	287.416.5-210.039	136.615.5-630.001	29.06.3-920.001

3. Inhibition of DNA synthesis

DNA synthesis was determined by incorporation of deuterium-labeled thymidine into the triacetic acid (TCA) precipitate of cultured PAP2 cells. PAP2 cells were plated at 3X 10/well⁴One cell was seeded in a 24-well plate for two days. Then, after preincubation with Compound 1 in ethanol, 0.5. mu. Ci of methyl- [3H ] was added]Thymidine (specific activity 82Ci/mmol) and labeled for 4 hours. The cells were then washed with 1ml of cold Phosphate Buffered Saline (PBS). Dissolved with 0.2ml of 4% sodium lauryl sulfate. The cell extract was precipitated with 1ml 30% TCA and stored in ice bath for 1 hour. The precipitate was collected by filtration on a glass fiber filter and washed with 5ml of 5% TCA. Radioactivity on the filter paper was counted using a liquid crystal scintillation counter.

TABLE 4

Inhibition of DNA Synthesis in Ras-transfected cells (PAP2) by Compound 1

Incorporation of 3H-thymidine into DNA (cpm)

Concentration of Compound 1
								0	0.5μM	2μM	5μM	10μM	25μM
cpmsem% change p	5302172960	456075354-140.427	371302590-300.075	302263945-430.016	187721481-650.009	141002766-730.005

The calculated IC50 value of compound 1 for DNA synthesis inhibition in PAP2 cells was 3.75 μ M.

The results in Table 4 show that compounds of formula (I), particularly Compound 1, are capable of inhibiting DNA synthesis in Ras-transfected cells. 4. Induction of apoptosis

It has now been shown that compounds of formula (I) have a selective effect on the promotion of cell death in the PAP2 cell line at concentrations of 5. mu.M or higher. This specific effect of selectively promoting apoptosis in cancer cells was demonstrated in the other two human cancer cell lines (HepG2 and SW480) treated with compound 1, indicating that these compounds not only inhibit cell growth but also produce destruction of the cancer cells themselves (table 5). This was confirmed by gel electrophoresis of the test blots showing DNA fragments and apoptosis.

TABLE 5

Effect of Compounds on cell viability in human cancer cell lines

Cell lines	Number of cells surviving at 0 hour	Number of cells surviving at 24 hours	% change
				SW480	38′800	0	-100.0％
HepG2	214′700	82 000	-61.8％

Cells were cultured in 24-well plates for 48 hours before treatment. 10 μ M compound treatment for 24 hours. Count: cells were trypsinized and viable cells were counted. 5. Effect of Compound 1 on a broad range of human cancer cell lines

The antiproliferative activity of compound 1 was tested in a wide range of established human-derived cancer cell lines to confirm the potential use of these compounds for human-related cancers (table 6). The results indicate that compound 1 is effective in reducing cell proliferation in more than 90% of the 50 human cancer cell lines tested. Therefore, it is believed that the compounds of formula (I) can be used to treat a wide range of human cancers with and without Ras mutations.

TABLE 6

Antiproliferative effect of Compound 1 on cell lines derived from human tumors

Tumor type	Cell lines	Inhibition of proliferation (% control)
			Leukemia (leukemia)	CCRF-CEM	-61.8
Leukemia (leukemia)	HL-60(tb)	-95.3
			Leukemia (leukemia)	K-562	-61.2
Non-small cell lung cancer	A549/ATCC	-55.0
			Non-small cell lung cancer	EKVX	-27.0
Non-small cell lung cancer	HOP-62	-13.8
			Non-small cell lung cancer	HOP-92	-85.1
Non-small cell lung cancer	NCl-H23	-21.8
			Non-small cell lung cancer	NCl-H322M	-12.5
Non-small cell lung cancer	NCl-H460	-40.7
			Non-small cell lung cancer	NCL-H522	-59.8

Cancer of colon	COLO 205	-55.9
			Cancer of colon	HCC-2998	-50.7
Cancer of colon	HCT-116	-68.7
			Cancer of colon	HCT-15	-19.5
Cancer of colon	HT29	-83.0
			Cancer of colon	KM12	-32.5
Cancer of colon	SW-620	-21.4
			CNS cancer	SF-268	-21.2
CNS cancer	SF-295	-49.6
			CNS cancer	SF-539	-1.5
CNS cancer	SNB-19	-55.8
			CNS cancer	SNB-75	-54.4
CNS cancer	U251	-41.6
			Melanoma (MEA)	LOX IMVI	-18.7
Melanoma (MEA)	MALME-3M	-20.4
			Melanoma (MEA)	M14	-66.7
Melanoma (MEA)	SK-MEL-2	-6.1
			Melanoma (MEA)	SK-MEL-28	-25.0
Melanoma (MEA)	SK-MEL-5	-51.8
			Melanoma (MEA)	UACC-257	-26.9
Melanoma (MEA)	UACC-62	-30.9
			Ovarian cancer	IGROV1	-24.4
Ovarian cancer	OVCAR-3	-47.3
			Ovarian cancer	OVCAR-4	-18.9
Ovarian cancer	OVCAR-5	-17.6
			Ovarian cancer	OVCAR-8	-37.6
Ovarian cancer	SK-OV-3	-28.1
			Renal cancer	786-0	-28.7
Renal cancer	ACHN	-43.4

Renal cancer	CAKI-1	-31.1
			Renal cancer	SN12C	-1.0
Renal cancer	TK-10	-18.6
			Renal cancer	UO-31	-35.9
Prostate cancer	PC-3	-75.9
			Prostate cancer	DU-145	-21.3
Breast cancer	MCF-7	-57.0
			Breast cancer	MCF-7/ADR-RES	-49.3
Breast cancer	MDA-MB-231/ATCC	-50.9
			Breast cancer	MDA-MB-435	-51.6
Breast cancer	MDA-N	-43.3
			Breast cancer	BT-549	-51.3
Breast cancer	T-47D	-62.1

EXAMPLE 2 in vivo results inhibition of tumor growth

On the day, 0.5ml of PAP2 cells (5X 10 cells per 0.5ml of DMEM) were injected subcutaneously (s.c.) into 7 to 9-week female nude mice (Swiss nu/nu). Treatment was performed orally with compound 1 (50mg/kg) on the same day. Compound 1 (0.03% w/w) was obtained mixed with food in the treatment group (n-18) and on the control group (n-22) on a diet without compound. On day 15, the mice were weighed, sacrificed and tumors removed and weighed.

TABLE 7a

Oral treatment (50mg/kg) with Compound 1 for subcutaneous injection

Effect of tumor weight production in nude mice with PAP2 cells

Animal group	Body weight (g) (mean. + -. sem)	Tumor weight (g) (mean. + -. sem)
			Control group (n ═ 21)	24.2±0.5	0.228±0.041
Treatment group (n ═ 18)	24.6±0.7	0.053±0.016

% change	+2	-77
			p	0.6470	0.0006

The results in table 7a show a significant decrease in mean tumor weight in the treated group, indicating that compound 1 is a potent anti-tumor agent. The weight of mice was the same in the control group and the treatment group, so compound 1 was determined to be non-toxic.

In another experiment, Compound 1 was tested at doses of 12.50 and 100 mg/kg. The results in table 7b show that compound 1 inhibits tumor growth at doses as low as 12.5mg/kg and inhibits tumor growth in nude mice in a dose-dependent manner. These results demonstrate that the compounds of formula (I), in particular compound 1, are potent orally active antitumor compounds and have no toxic effect on normal cells, tissues or organs.

TABLE 7b

For oral therapeutic injection with different doses of Compound 1

PAP2 cells in nude mice observed tumor weight reduction

Animal group	Body weight (g) (mean. + -. sem)	Tumor weight (g) (mean. + -. sem)
			Control group (n ═ 14)	24.7±0.6	0.276±0.056
Treatment group, 12.5mg/kg (n ═ 5)% change p	23.9±0.8-30.4562	0.053±0.026-810.0472
			Treatment group, 50mg/kg (n ═ 6)% change p	27.2±1.0+100.0523	0.037±0.020-870.0211
Treatment group100mg/kg (n ═ 6)% change p	24.5±0.3-10.7619	0.025±0.011-910.0151

Example A2- (3, 5-diisopropyl-4-hydroxyphenyl) -ethenylene-1, 1-bisphosphate tetraisopropyl ester

Titanium tetrachloride (13.83g, 73mmol) was added dropwise to anhydrous THF (80ml) and held at 0 ℃. The resulting mixture was treated with 3, 5-diisopropyl-4-hydroxybenzaldehyde (5.0g, 24mmol), tetraisopropyl methylenediphosphonate (10.85g, 32mmol) and N-methylmorpholine (14.71g, 146mmol) in that order at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours and 80ml of water were added. The quenched reaction mixture was extracted with diethyl ether (3 × 60ml) and the combined ether fractions were extracted with saturated NaCl solution until the aqueous solution was washed to neutral pH. Over MgSO₄After drying, the organic solvent is evaporated and washed with CH on silica gel₂Cl₂Column chromatography of the residue with a mixture of MeOH (95: 5) as eluent gave 8g (62%) of a solid; mp is 87-88 ℃. MS: 532/e: m⁺：367：M⁺-PO₃iPr₂NMR(CDCl₃) δ 8.22(dd, J31 and 48Hz, 1H): Ph-CH ═ C-P₂

7.7(s, 2H): the aromatic substance is an aromatic substance having a high degree of aromaticity,

5.6(s，1H)：OH，

4.85-4.63(2m，4H)：P-O-CHMe₂，

3.16 (detached, 2H): Ph-CHMe₂，

1.39, 1.36, 1.27, 1.23 and 1.16(8d, 36H total): P-O-CHMe₂And Ph-CHMe₂。

Example B2- (3, 5-diisopropyl-4-hydroxyphenyl) -ethylene-1, 1-bisphosphate tetraisopropyl ester

Tetraisopropyl 2- (3, 5-diisopropyl-4-hydroxyphenyl) -vinylene-1, 1-diphosphate (5g, 9.4mmol) is dissolved in ethanol (50ml) and the solution is hydrogenated at room temperature at 50psi over 2g 10% palladium on carbon for 4 hours. The crystals are filtered, the solvent is evaporated and washed with CH on silica gel₂Cl₂Column chromatography of the residue with a mixture of MeOH (95: 5) as eluent gave 2.5g (50%) of a solid; mp is 90-91 ℃. MS: 534, m/e: m⁺，369：M⁺-PO₃iPr₂NMR(CDCl₃) δ 6.94(m, 2H): the aromatic substance is an aromatic substance having a high degree of aromaticity,

4.8-4.7(m，5H)：P-O-CHMe₂and an OH group, and a nitrogen-containing compound,

3.2-3.1 (several m, total 6H): Ph-CH₂-CH and Ph-CHMe₂，

2.51(tt, J ═ 6 and 24H, 1H): Ph-CH₂-CH，

1.33-1.21 (several d, total 36H): P-O-CHMe₂And Ph-CHMe₂。

Example C

Treatment of 3, 4, 5-trimethoxybenzaldehyde (7g, 35.7mmol) with titanium tetrachloride, tetraisopropyl methylenediphosphonate and N-methylmorpholine in THF as described in example A gave 11.5g (62%) of the title compound. MS: 522 for m/e: m⁺：357：M⁺-PO₃iPr₂NMR(CDCl₃) δ 8.21(dd, J30 and 48Hz, 1H): Ph-CH ═ C-P₂

7.28(s, 2H): the aromatic substance is an aromatic substance having a high degree of aromaticity,

4.85-4.63(2m，4H)：P-O-CHMe₂，

3.9(t，9H)：Ph-OMe，

1.4, 1.36 and 1.22(4d, 24H total): P-O-CHMe₂. Example D2- (3, 4, 5-trimethoxyphenyl) -ethylene-1,1-Tetraisopropyl diphosphate

Hydrogenation of 2- (3, 4, 5-trimethoxyphenyl) -ethenylene-1, 1-diphosphonic acid tetraisopropyl ester (7g, 13.4mmol) at 10% Pd/C as described in example B gave 5.7g (81%) of the title compound. MS: m/e 524: m⁺，359：M⁺-PO₃iPr₂NMR(CDCl₃) δ 6.55(s, 2H): the aromatic substance is an aromatic substance having a high degree of aromaticity,

4.8-4.7(m，4H)：P-O-CHMe₂，

3.85 and 3.81(2s, 9H): the Ph-OMe is a function of Ph-OMe,

3.17(dt, J ═ 6 and 16Hz, 2H): Ph-CH₂-CH，

2.50(tt, J ═ 6 and 24Hz, 1H): Ph-CH₂-CH，

1.33-1.26 (several d, 24H total): P-O-CHMe₂。

Example E2- (3-tert-butyl-4-hydroxy-5-methylphenyl) -ethenylene-1, 1-diphosphonic acid tetraisopropyl ester

Treatment of 3-tert-butyl-4-hydroxy-5-methylbenzaldehyde (6g, 31.3mmol) with titanium tetrachloride, tetraisopropyl methylenediphosphonate and N-methylmorpholine in THF as described in example A gave 4.2g (62%) of the title compound. MS: m/e 518: m⁺：353：M⁺-PO₃iPr₂NMR(CDCl₃) δ 8.19(dd, J30 and 48Hz, 1H): Ph-CH ═ C-P₂

7.71-7.76(m, 2H): the aromatic substance is an aromatic substance having a high degree of aromaticity,

5.6(s，1H)：OH，

4.8-4.7(2m，4H)：P-O-CHMe₂，

2.26(s，3H)：Ph-Me，

1.40(s，9H)：Ph-t-Bu，

1.38, 1.36, 1.24 and 1.20(8d, 24H total): P-O-CHMe₂。

Example F2- (3-tert-butyl-4-hydroxy-5-methylphenyl) -ethylene-1, 1-bisphosphate tetraisopropyl ester

Hydrogenation of 2- (3-tert-butyl-4-hydroxy-5-methylphenyl) -ethenylene-1, 1-diphospho tetraisopropyl ester (5g, 9.6mmol) over 10% Pd/C as described in example B gave 2.9g (58%) of the title compound. MS: m/e 520: m⁺：355：M⁺-PO₃iPr₂NMR(CDCl₃) δ 7.02 and 6.92(2m, 2H): the aromatic substance is an aromatic substance having a high degree of aromaticity,

4.8-4.7(m，5H)：P-O-CHMe₂and an OH group, and a nitrogen-containing compound,

3.11(dt, J ═ 6 and 17Hz, 2H): Ph-CH₂-CH，

2.49(tt, J ═ 6 and 24Hz, 1H): Ph-CH₂-CH，

2.21(s，3H)：Ph-Me，

1.39(s，9H)：Ph-t-Bu，

1.31, 1.26 and 1.24(3d, 24H): P-O-CHMe₂。

Example G2- (3-ethoxy-4-hydroxyphenyl) -ethenylene-1, 1-bisphosphate tetraisopropyl ester

3-ethoxy-4-hydroxybenzaldehyde (6g, 36.1mmol) was treated with titanium tetrachloride, tetraisopropyl methylenediphosphonate and N-methylmorpholine in THF as described in example A to give 4.2g (62%) of the title compound. MS: 492, m/e: m⁺：327：M⁺-PO₃iPr₂NMR(CDCl₃) δ 8.19(dd, J31 and 48Hz, 1H): Ph-CH ═ C-P₂

7.9, 7.3 and 6.91(3m, 3H): the aromatic substance is an aromatic substance having a high degree of aromaticity,

6.2(s，1H)：OH，

4.85-4.63(2m，4H)：P-O-CHMe₂，

4.19(q，J＝7Hz)：Ph-OCH₂-CH₃，

1.46(t，J＝7Hz)：Ph-OCH₂-CH₃，

1.39, 1.36, 1.23 and 1.21(4d, 24H total): P-O-CHMe₂。

Example H2- (3-ethoxy-4-hydroxyphenyl) -ethylene-1, 1-bisphosphate tetraisopropyl ester

Hydrogenation of 2- (3-ethoxy-4-hydroxyphenyl) -ethenylene-1, 1-diphospho tetraisopropyl ester (5g, 10.16mmol) over 10% Pd/C as described in example B gave 4.5g (90%) of the title compound. MS: 494 of m/e: m⁺：329：M⁺-PO₃iPr₂NMR(CDCl₃) δ 6.85 and 6.75 (several m, 3H): the aromatic substance is an aromatic substance having a high degree of aromaticity,

5.65(s，1H)：OH，

4.8-4.7(m，4H)：P-O-CHMe₂，

4.10(q，J＝7Hz)：Ph-OCH₂-CH₃，

3.14(dt, J ═ 6 and 16Hz, 2H): Ph-CH₂-CH，

2.45(tt, J ═ 6 and 24Hz, 1H): Ph-CH₂-CH，

1.43(t，J＝7Hz)：Ph-OCH-CH₃，

1.32-1.25(4 partially overlapping d, 24H total): P-O-CHMe₂。

EXAMPLE I2- (3, 5-di-tert-butyl-4-methoxyphenyl) -ethenylene-1, 1-bisphosphate tetraisopropyl ester

3, 5-di-tert-butyl-4-methoxybenzaldehyde (2.0g, 8.1mmol) was treated with titanium tetrachloride, tetraisopropyl methylenediphosphonate and N-methylmorpholine in THF as described in example A to give 3.0g (72%) of the title compound. mp-66-67 ℃ MS: m/e 518: m⁺：381：M⁺-PO₃Et₂NMR(CDCl₃) δ 8.26(dd, J30 and 48Hz, 1H): Ph-CH ═ C-P₂

7.8(s, 2H): the aromatic substance is an aromatic substance having a high degree of aromaticity,

4.25-4.00(2m，8H)：P-O-CH₂-CH₃，

3.69(s，3H)：Ph-OMe.

1.44(s，18H)：Ph-t-Bu，

1.38 and 1.70(2t, 12H): Ph-OCH₂-CH₃. Example J1- (3, 5-di-tert-butyl-4-hydroxyphenyl) -butylidene-2, 2-bisphosphate tetraisopropyl ester

Tetraisopropyl trimethylene-1, 1-diphosphate is prepared by reacting tetraisopropyl methylene diphosphate with 3 equivalents of iodoethane in THF in the presence of NaH.

Tetraisopropyl propylene-1, 1-diphosphate (2.2g, 6.0mmol) is added to a suspension of 60% NaH (0.5g, 12.0mmol) in 20ml of anhydrous THF and the mixture is stirred until the NaH disappears. 3, 5-di-tert-butyl-4-hydroxybenzyl chloride (1.5g, 6mmol) in 10ml THF was added and the mixture was refluxed overnight. After completion, CHCl was applied to silica gel₃Column chromatography with AcOEt (8: 2) as eluent gave 1.5g (42%) of the title compound. MS: 590 m/e: m⁺：425：M⁺-PO₃Pr₂Base peak 341: m⁺-2×propenmp＝131-132℃

Example K typical active ingredients of the compound formulation: compound 1 inactive ingredients: pregelatinized starch NF3 opaque dark blue gelatin capsule

Representative formulations

Typically a batch of 2000 capsules

Components	g/batch
				1mg	10mg	50mg
	Compound 1	2.0	20.0				100.0
Pregelatinized starch NF				431.6	408.8	291.0
	In all	433.6	428.8				391.0

Claims (16)

1. Use of a phenol-substituted gem-diphosphate in the preparation of a medicament for the treatment or prophylaxis of a tumor disease, wherein the phenol-substituted gem-diphosphate is selected from the group of compounds of the general formula (I),wherein: -Z¹、Z²、Z³And Z⁴Are identical OR different and are-OR, where R is H, a linear, branched OR cyclic alkyl radical having 1 to 8 carbon atoms, -OM, where M is a cation, -NR₂Wherein R has the same meaning as defined above, -Z¹、Z²And Z³、Z⁴May form an alkylene dioxycycle having from 2 to 8 carbon atoms, -X¹、X²Are identical or different and are H, a halogen atom, a linear, branched or cyclic alkyl or alkoxy radical having 1 to 8 carbon atoms, X³Is H, an alkyl radical R having from 1 to 4 carbon atoms¹Acyl C (O) R¹Carbamoyl C (O) NHR¹Wherein R is¹As described above, X³O and two other substituents X¹Or X²One of which may form an alkylenedioxy ring containing from 1 to 4 carbon atoms, -A is-CH ═ CH-CH₂-、-(CH₂)_n-、-O(CH₂)_n-、-S-、-SO₂-、-S(CH₂)_n-、-SO₂(CH₂)_n-, where n is an integer from 1 to 7, or A and B form a compound of the formula- (CH ═ CH)_k-(CH₂)_d-CH ═ alkylene, where k is 0 or 1 and d is an integer from 0 to 4, -B is H, an alkyl group containing 1 to 4 carbon atoms, -t is 0 or 1, with the proviso that when a is- (CH ═ CH)_k-(CH₂)_d-CH ═ where k and d are as described above, when t is 0.

2. Use according to claim 1, wherein the neoplastic disease is selected from the group consisting of cancers of the hematopoietic and immune systems, including lymphomas and leukemias, and pancreatic, colon, breast, thyroid, brain, lung, head and neck cancers.

3. Use according to claim 1 of a diphosphate selected from among the compounds of formula [ I ] for the preparation of a medicament for the prevention of metastatic invasion of normal cells by cancer cells.

4. Use according to any one of claims 1 to 3, the phenol-substituted alkylene diphosphate being selected from the compounds of formula (Ia)Wherein X¹、X²、X³、A、B、Z¹、Z²、Z³And Z⁴As defined in claim 1.

5. Use according to any one of claims 1 to 3, of a phenol-substituted alkenylene diphosphate selected from the compounds of formula (Ib)Wherein X¹、X²、X³、k、d、Z¹、Z²、Z³And Z⁴As defined in claim 1.

6. Use according to any one of the preceding claims, wherein Z is¹、Z²、Z³And Z⁴Are identical or different and are selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl and tert-butyl.

7. Use according to claim 6, wherein Z¹、Z²、Z³And Z⁴Are the same.

8. Use according to claim 7, wherein Z¹、Z²、Z³And Z⁴Is isopropyl.

9. Use according to any one of the preceding claims, wherein X¹And X²Are identical or different and are selected from linear or branched alkyl and alkoxy groups having from 1 to 5 carbon atoms.

10. Use according to claim 9, wherein X¹And X²Are identical or different and are selected from the group consisting of methyl, ethyl-n-propyl, sec-butyl, tert-butyl, methoxy and ethoxy.

11. Use according to claim 10, wherein X¹And X²Are the same.

Use according to claim 1, wherein X¹And X²Are both tert-butyl groups.

13. Use according to any one of the preceding claims, wherein X³Selected from hydrogen, C_1-4Alkyl and C_1-4An alkanoyl group.

14. Use according to claim 13, wherein X³Is a hydrogen atom.

15. Use according to any one of claims 1 to 3, wherein the compound of formula (I) is tetraisopropyl 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -ethylene-1, 1-diphosphate.

16. Use according to any one of claims 1 to 3, wherein the phenol-substituted gem-diphosphate is selected from the group consisting of: 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -ethylene-1, 1-bisphosphate Tetramethyl 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -ethylene-1, 1-bisphosphate tetraethyl 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -ethylene-1, 1-bisphosphate tetra-n-propyl 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -ethylene-1, 1-bisphosphate tetra-n-butyl 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -ethylene-1, tetraisopropyl 1-diphosphate 2- (3, 5-di-sec-butyl-4-hydroxyphenyl) -ethylene-1, 1-tetraisopropyl diphosphate 2- (3, 5-diisopropyl-4-hydroxyphenyl) -ethylene-1, 1-tetraethyl diphosphate 2- (3, 5-diisopropyl-4-hydroxyphenyl) -ethylene-1, 1-tetraisopropyl diphosphate 2- (3-tert-butyl-4-hydroxy-5-methylphenyl) -ethylene-1, 1-tetraethyl diphosphate 2- (3-tert-butyl-4-hydroxy-5-methylphenyl) -ethylene-1, 1-tetraisopropyl diphosphate 3, 5-di-tert-butyl-4-hydroxy-phenyl-thiomethylene-1, 1-diphospho tetraethyl ester 3, 5-di-tert-butyl-4-hydroxy-phenyl-thiomethylene-1, 1-diphospho tetraisopropyl ester 2- (3, 4, 5-trimethoxyphenyl) -ethylene-1, 1-diphospho tetraisopropyl ester 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -ethylene-1, 1-diphospho dibutyl diethyl ester 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -ethylene-1, 1-diphospho diethyldiisopropyl ester 1- (3, 5-di-tert-butyl-4-hydroxyphenyl) -butylene-2, tetraethyl 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -ethylene-1, 1-bis (2-oxo-1, 3, 2-dioxaphospholane) 1- (3, 5-di-tert-butyl-4-hydroxyphenyl) -butylene-2, 2-tetraisopropyl diphosphate 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -vinylene-1, 1-tetramethyl diphosphate 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -vinylene-1, 1-tetraethyl diphosphate 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -vinylene-1, 1-bisphosphate tetraisopropyl 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -vinylene-1, 1-bisphosphate tetra-n-propyl 2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -vinylene-1, 1-bisphosphate tetra-n-butyl 2- (3, 5-di-sec-butyl-4-hydroxyphenyl) -vinylene-1, 1-bisphosphate tetraethyl 2- (3, 5-di-sec-butyl-4-hydroxyphenyl) -vinylene-1, 1-bisphosphate tetraisopropyl 2- (3, 5-diisopropyl-4-hydroxyphenyl) -vinylene-1, tetraisopropyl 1-diphosphate 2- (3-tert-butyl-4-hydroxy-5-methylphenyl) -ethenylene-1, 1-diphosphonate tetraethyl 2- (3-tert-butyl-4-hydroxy-5-methylphenyl) -ethenylene-1, 1-diphosphonate tetraisopropyl 2- (3, 5-dimethoxy-4-hydroxyphenyl) -ethenylene-1, 1-diphosphonate tetraethyl 2- (3, 5-dimethoxy-4-hydroxyphenyl) -ethenylene-1, 1-diphosphonate tetraisopropyl 2- (3, 4, 5-trimethoxyphenyl) -ethenylene-1, 1-diphosphonate tetraisopropyl 2- (3-ethoxy-4-hydroxyphenyl) Tetraisopropyl (4- (3, 5-di-tert-butyl-4-hydroxyphenyl) -1, 3-butadienylene-1, 1-bisphosphate Tetraisopropyl (2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -vinylene-1, 1-bisphosphate Dibutyldiethyl (2- (3, 5-di-tert-butyl-4-hydroxyphenyl) -vinylene-1, 1-bisphosphate Diethyldiisopropyl (2- (3, 5-di-tert-butyl-4-methoxyphenyl) -vinylene-1, tetraethyl 1-diphosphate 2- (3, 4-methylenedioxyphenyl) -vinylene-1, 1-diphosphonate and tetraethyl 2- (3, 4-ethylenedioxyphenyl) -vinylene-1, 1-diphosphonate.