DE4431175A1 - New drugs containing chelating agents - Google Patents

Abstract

The invention concerns novel drugs which contain chelate-forming agents and essential amino acids or their derivatives which are optionally complexed with bivalent metal ions. The novel drugs further contain bivalent elements or compounds of bivalent elements.

Description

The invention relates to new drugs, the chelating agents as well as essential amino acids or derivatives thereof which optionally complexed with divalent metal ions and divalent elements or compounds of divalent Elements included.

Chelating agents have long been successful as Medicines or components of medicines in the Medicine has been used or at least in the discussion, for example in the therapy of metal ion poisoning or in the inhibition of enzymes by binding Cofactors. It is known that the steric Conformation of the chelating agents and their ability to Receptor binding to a large extent the therapeutic Affect effects. Many of the prior art known chelating agents, however, induce side effects, that oppose a broad medical application (see e.g. Mazzarello et al., Schwe. Arch. Neurol. Psych. 131: 175-179 (1982)). The furthest so far  widespread chelating agent used as a drug Azyclovir, which has already had considerable success in the medical practice (e.g. Corey et al., New Engl. J. Med. 306 (1982), 1313). However, this is also Medicines do not work well for all indications applicable. For example, it is known that azyclovir in AIDS therapy can cause significant side effects can. Another known in the art The drug is RODILEMID®, which is an antiviral, anti-inflammatory, immunomodulatory and neurotrophic Has effects. This medicine has been found in the Past particularly due to its good tolerability and also recommended due to its good therapeutic effect; cf., e.g. B. Dinu and Dinu, La Sante Publique (1985), 3-30. However, there is a need for additional drugs that the therapeutic properties of the above Have chelating agents, but beyond have better / different tolerance spectra and one have another / different application.

In many physiological processes, u. a. in connection play with the above-mentioned clinical pictures Trace elements, e.g. B. zinc, an important role. Changes trace element concentrations (oligo element Modulations) can be very useful in treatment, even if the exact mechanism of action of the oligo element Modulation has not yet been fully clarified.

The object of the present invention was thus pharmaceuticals with the positive properties described above to provide that in a greater number Indications or other indications without harmful Side effects can be used and thereby u. a. as a chelating agent and / or oligo element Modulators can act. This task is carried out by the in the embodiments described solved.  

The invention thus relates to a medicament containing

• (a) a polyaminopolycarboxylic acid or a derivative or a complex of them;
• (b) an essential amino acid or a derivative thereof; and
• (c) a divalent element or a combination of one divalent element; or
• (a ') a polyaminopolycarboxylic acid or a derivative or a complex of them; and
• (b ′) a complex of an essential amino acid and divalent metal ions;

with the exception of the combination

• (a) EDTA-CaNa₂
• (b) cysteine in a concentration of 0.38-2.3% by weight based on the sum of the three components
• (c) calcium gluconate;

and optionally a pharmaceutically acceptable one Carrier.

The pharmaceuticals according to the invention are characterized by a with administration of usual concentrations with optimal Effects of lack of toxicity. Furthermore is for an already known medicine with a similar composition that in the present invention however as such is not claimed and the name RODILEMID® has been shown to be a longer one Administration has no mutagenic potential. (Moralidhara et al., Food Chem. Toxicol. December 12, 1991, 845-849). It it is believed that the mechanism of action of the fiction medicines can be of different types. For example, the antiviral effectiveness of the Medicament according to the invention are based on the fact that Withdrawal of zinc ions by the chelating agent disrupts the Zinc finger function with various retroviral proteins brings itself. The amino acid component could be in the Embodiment as a significant regulatory cysteine Role between T cells and macrophages and thus  develop immunomodulatory effects. In any case there may also be synergistic effects of the individual Components of the pharmaceuticals according to the invention come that these for use in various therapies predestine.

In a preferred embodiment of the invention Medicament are those mentioned in (a) and (a ′) above Polyaminopolycarboxylic acids EDTA, DTPH, DCTA or derivatives or complexes, preferably salts thereof, preferably EDTA- CaNa₂.

In a further preferred embodiment of the The medicament according to the invention is that in (b) and (b ′) called amino acid cysteine, histidine or methionine and that Derived from a precursor of cysteine or histidine or of methionine or N-acetylcysteine, N-acetylhistidine or N-acetylmethionine.

In a further preferred embodiment of the The medicament according to the invention is that mentioned in (c) divalent element calcium and the derivative thereof Calcium salt, preferably calcium lactate, calcium gluconate or calcium levulinate.

In a further preferred embodiment, the in (b ′) called complex

• (i) cysteine and copper;
• (ii) serine and copper;
• (iii) tyrosine and copper;
• (iv) cysteine and zinc;
• (v) serine and zinc; or
• (vi) tyrosine and zinc.

In a further preferred embodiment, the Medicaments according to the invention at least one of the following Mixtures

• 1. EDTA-CaNa₂, N-acetylcysteine and calcium gluconate;
• 2. EDTA-CaNa₂, calcium gluconate and cysteine in one Concentration greater than 2.3 wt .-%, based on the sum of the three components;
• 3. EDTA-CaNa₂ and a complex of cysteine and copper;
• 4. EDTA-CaNa₂ and a complex of serine and copper;
• 5. EDTA-CaNa₂ and a complex of tyrosine and copper;
• 6. EDTA-CaNa₂ and a complex of cysteine and zinc;
• 7. EDTA-CaNa₂ and a complex of serine and zinc; or
• 8. EDTA-CaNa₂ and a complex of tyrosine and zinc.

In a further preferred embodiment of the The medicament according to the invention comes in (a) and (a ′) called polyaminopolycarboxylic acid or the derivative or the Complex thereof in a concentration of 90-95% by weight, the essential amino acid or the derivative mentioned in (b) of which in a concentration of 0.3-3% by weight, the divalent element or the compound mentioned in (c) of a divalent element in a concentration of 3-7 % By weight, and the complex mentioned in (b ′) in a concentration of 3.3-7% by weight.

The above percentages are in each case on the sum of the components (a), (b) and (c) or (a ′) and (b ′) based. The invention further relates to the use of Medicament according to the invention for the treatment of especially viral, preferably retroviral Diseases.

As mentioned above, it is believed that the antiviral mechanism of action on one by that Medicament according to the invention induced disorder of viral metal ion availability is based. Especially at Retroviruses are known to have very strong genomic RNA Binds nucleocapsid proteins, causing the nucleocapsid Structure is formed. The so-called NC protein is used  needed for RNA dimerization, encapsulation and to initiate reverse transcription in human Retroviruses. The NC7p7 protein of the HIV-1 nucleocapsid is necessary for the formation of infectious particles and contains two Zinc finger of the Cys-X2-Cys-X4-His-X4-Cys type. Probably causes the medicament according to the invention that this Zinc fingers their biological capabilities either through a steric change in the binding structure or by Loss of zinc ions lose, in any case the three-dimensional structure of the nucleotide sequence changed which will ultimately be the loss of the three-dimensional binding activity and thus loss that brings infectiousness with it.

The invention further relates to the use of Medicament according to the invention and a medicament containing the above mentioned disclaimer Composition for the treatment of AIDS.

The invention further relates to the use of the Medicament according to the invention for the treatment of Inflammation, preferably from polyradiculonevritis.

The invention further relates to the use of a Medicament according to the invention as an immunomodulator; e.g. B. Immune stimulation in flu and paragrippals Infections.

The invention also relates to the use of a Medicament according to the invention for the treatment of Autoimmune diseases.

In a particularly preferred embodiment, the Invention the use of the medicament according to the invention for the treatment of multiple sclerosis, the Guillaume-Barre- Syndrome, as well as herpes simplex, herpes zoster and facial Paresis.  

In a further preferred embodiment, the Invention the use of an inventive Medicinal product for the treatment of neurodegenerative diseases.

Finally, the invention relates to the use of a Medicament according to the invention and a medicament, the composition mentioned in the above disclaimer has, as an anti-cancer agent.

Fig. 1 shows the effectiveness of the medicament according to the invention.

The examples illustrate the invention.

Example 1: Inhibition of HIV-1 Infections by EDTA CaNa₂ cysteine calcium gluconate (RODILEMID®)

The anti-HIV-1 activity and cytotoxicity of EDTA CaNa₂- Cysteine calcium gluconate was tested in the MT-4 cell test (Harada et al., J. Immunol. Meth. 92 (1986), 177-181) and the results with the toxic and inactive anti-HIV-1 Concentrations of four RODILEMID® derivatives (lAlH, lA09, lA2N, lAlN), DMSO / H₂O (blind test, M) and azidothymidine (3'-Azido-2 ', 3'-dideoxythymidine, AZT) compared.

The infection of the MT-4 cells became the IIIB strain of HIV 1 virus (Popovic et al., Science 224 (1984), 497-500) in CEM cells propagated. MT-4 and CEM cells (freely available, e.g. B. from ATCC) were in RPMI-1640-medium (Gibco, Karlsruhe, Germany), which with 20 vol .-% heat inactivated fetal calf serum (FCS, Seromed, Berlin, Germany) and Antibiotics had been added in the usual concentration. During the growth of the cells for the MT-4 cell test this in RPMI-1640 medium with 25 nM HEPES (2-4 (2-hydroxy ethyl) -1-piperazinylethanesulphonate) and FCS.  

The MT-4 cell test was carried out in microtiter plates with 96 Deepening carried out. For measuring the Cyto Pathogenicity of the active substances were serial dilutions of RODILEMID® with 3 × 10⁴ MT-4 cells per well incubated. The anti-HIV-1 activity was in the presence of the Virus measured. The final virus concentrations were 100 TCID₅₀, with one unit TCID₅₀ as the infectious dose is defined in the cell culture, which is the number of living Cells reduced by 50%. Three days after the infection was over fresh medium added to each well. Four days after of infection were 0.1 µCi 3 H-thymidine per well (Amersham, Braunschweig, Germany) added. After 20 The cellular DNA was held on glass fiber filters for hours harvested and the incorporated ³HThymidine in one Scintillation counter using PPO, POPOP and Toluene as a scintillator (Roth, Karlsruhe, Germany) certainly. The antiviral activity and the cytotoxic Concentration of RODILEMID® was associated with the activities and Concentrations of the four RODILEMID® from the blind test M and compared to that of AZT.

For this purpose, the ³H-thymidine incorporation in MT-4 cell DNA was applied in the presence or absence of HIV-1 against various concentrations of RQDILEMID® and AZT (cf. FIG. 1). The minimum toxic and maximum anti-HIV-1 concentrations of these compositions are shown in Table 1.

RODILEMID® was toxic to the MT-4 cells at a concentration of 1250 µg / ml ( Fig. 1a, dashed line). A RODILEMID® concentration of 625 µg / ml had no influence on the cell growth and at this concentration a strong anti-HIV-1 activity could be determined via a strong ³H-thymidine incorporation ( Fig. 1a, solid line). Anti-HIV-1 activity of RODILEMID® could be determined at concentrations between 625 µg / ml and 39 µg / ml.

AZT was toxic to those in MT-4 cells at a concentration of 250 µM ( Fig. 1b, dashed line). The anti-HIV-1 activity of AZT could be measured in the range of 25000 nM and 25 nM ( FIG. 1b, solid line). A measurable difference in the anti-HIV-1 activity of RODILEMID® and the derivatives could not be determined. In terms of toxicity, only lAlH is less toxic to MT-4 cells than RODILEMID®: MT-4 cells died at a concentration of 2500 µg / ml lAlH. The three other derivatives lA09, lA2N and lAlN were toxic in the same range as RODILEMID® (1250 µg / ml).

The selectivity index SI (see Table 1) defined as 100% Cell toxicity concentration divided by 100% antiviral Concentrations for a compound / composition was for AZT highest (10,000). So RODILEMID® was with an SI out of 32 about 300 times less active than AZT and twice less active than the RODILEMID® derivative lAlH.  

Table 1

Toxicity and anti-HIV activities of RODILEMID® derivatives and non-RODILEMID® derivatives

Example 2 Treatment regimen for the use of the Medicament according to the invention for MS, AIDS, herpes simplex, Herpes zoster and Guillaume-Barre syndrome

The treatment process consists of long-term treatment with the drug and depends on the development of the clinical symptoms of the patient and other usual para-clinical parameters.

The treatment regimen includes active periods that coincide with Alternate periods when the drug is not is used. The treatment is longer through a phase alternating periods and a phase of short  alternating periods marked. The daily doses vary between 7 and 70 mg per kg of body weight as intramuscular, intravenous injections or as an oral dose be administered. Unless oral therapy is used the dose described above can be doubled will. After a break of 20 to 25 days, the Therapy resumed cyclically. After therapy cycles from 12 to 15 days a longer break from for example, insert three months, the length of which again from the clinical status and the usual para clinical parameters.

Therapy begins with the appearance of the patient to be treated Disease symptoms, in multiple sclerosis of a relapse, 15 to 70 mg of the invention according to the invention for 15 to 30 days Drug per kilogram of body weight as intramuscular or intravenous injections. Then treatment with doses of 7 to 35 mg Medicines per kilogram of body weight for 3 to 6 months continued, the Ver delivery routes can be used. Depending on The patient's clinical picture and the usual paraclinic Treatment can be parameterized after one to six months can be repeated again.

A strict diet must be followed during treatment whose job it is to increase the effectiveness of the Increase drug. From the one discussed above Mechanism of action and goes from pharmacological studies shows that the exogenous supply of divalent metal ions the biological effectiveness of the drug greatly reduced. Therefore, the following foods must be consumed during the active treatment phase can be avoided in any case: Grapefruit, lemons, oranges, cabbage, salads, leafy greens, Milk derivatives, cheese, yogurt, mineral water, grapefruit juice, Lemon juice, orange juice, beans, peas, soy, mushrooms, Tomatoes, carrots, beetroot, radishes, celery, cucumber,  Onions, wheat derivatives, milk-based pastries, liver, Kidney.

In contrast, apples, pears, bananas, cherries, peaches, Watermelons, grapes, coffee, tea, cereals, potatoes, White beets, eggs, veal, poultry, fish, fat, Rye bread, wheat bread, wheat starch, wheat, oatmeal, paddy rice, husked rice, rice starch, potato starch, flour to be consumed.

Example 3: RODILEMID® as an AIDS therapeutic

First, the toxic activity of RODILEMID® determined: In rats the LD₅₀ is 4790 mg / kg Body weight, in mice even 5940 mg / kg body weight.

Chronic toxicity due to intramuscular injections in Rats and beagle dogs showed that up to doses of 350 mg / kg body weight no toxicity, no mutagenic and no embryotoxic effects occur. This dose is 20 times higher than the therapeutically effective dose.

These results confirm the good application possibilities from RODILEMID® for use as a medicine in humans, for example in AIDS therapy.

Clinical trial

The first stage of therapy included eight AIDS patients clinically and immunologically checked. In addition, the Abbot kit used. All samples were redone either with the indirect or competitive Wellcozyme or the Abbot kit and subsequently tested by Western blot. The Patients consistently showed a T- at the start of the studies Cells number of about 450 per mm³. The patients were only inpatient for taking samples to the clinic convened. The immunological monitoring was carried out by the  Total lymphocyte and T₄- and T₈- counting Subpopulations using the rosette method supervised.

The RODILEMID® therapy started on the first day of immunological examinations by an intramuscular Injection per day over a period of 10 days. To The patients received one oral dose per month Day, 14 days a month. Four months later she had to Therapy should be stopped first. She was with afterwards 10 AIDS patients continued, seven of whom were the first examined group included and three new patients came along. These patients were given 10 doses per month administered, given only one dose per day has been. The immunological tests were carried out once per 45 days after restart of treatment and clinical monitoring lasted 30 and 18 months (for the three new patients) since the beginning of the Study.

In parallel to these studies, a group of seven HIV-1 infected patients aged 1 to 4.5 years had previously been identified as such, immunologically and clinically after starting oral therapy RODILEMID® observed. The therapy included the ver Dispensing 10 ampoules per month, one per day Ampoule was administered.

10 AIDS patients were observed as a control group. Your immunological development was simultaneous with that of RODILEMID® treated group examined. Beside that four children between the ages of 1 and 1.5 years who have HIV-1 had been transmitted horizontally for 18 months Administration of RODILEMID® observed. The children were One ampoule per day for 10 days. While of active RODILEMID® therapy lasting 10 days per month  any calcium intake, including through Dairy products, prevented.

In the group with eight AIDS patients was followed up four-month treatment with RODILEMID®, with a series of 10 intramuscular injections started and with oral Administration continued, a significant increase the total number of lymphocytes observed and an increase of all examined T-lymphocyte subpopulations. Here the T₄ lymphocyte population rose to one average value of 1000 per mm². At the same time, a improved clinical appearance of the patients observed.

As in the case of the already known indications for the Treatment with RODILEMID®, HIV, herpes zoster and viral Polyneuritis, the development of T₈ subpopulations continues the expected course and grows in the course of treatment. There is also an increase in T₄ lymphocyte subpopulations and a significant drop in IgG antibody from one Average from 3200 mg% to 1680 mg% after 5 months RODILEMID® treatment observed. During the 10th Months of the second phase of the RODILEMID®- Therapy was performed in all 10 patients receiving RODILEMID® orally was administered, d. H. the seven patients from the first Group and the three patients who joined later Increase in total lymphocyte count in all subpopulations observed. However, this increase was less significant than during the first phase, when the drug was intra was administered muscularly. This is probably due to that slightly lower dose. Oral after 3 months RODILEMID® treatment was the average value for that T₄ subpopulations increased from 408 / mm³ to 640 / mm³.

After the forced termination after 10 months after the first Therapy phase, one patient died 4 months after termination of the Treatment. She had been released from the hospital  after first significant improvements in the clinical picture had been observed.

After resuming treatment with the 10 AIDS Patients stopped one of the patients after taking four administration series of 10 ampoules each treatment and died 10 months later. At the time of giving up the Treatment she had a good clinical and immuno logical appearance. The remaining nine patients currently have a good clinical appearance. From a clinical point of view, the nine were Seventh stage patients on the Walter Reed (WR) scale. After 30 months of clinical monitoring for the seven Patients and 18 months for the three added Patients show this an advance from the ur initial assessment as WR 4 stage to WR 2 stage after the end of treatment.

The following are general clinical observations: The Adenopathies are fluctuating. The flare up of Herpes infections have been observed very rarely and only occurred still with reduced strength. Herpes zoster infections were healed within six to seven days Patients responded well to both specific and complementary treatments against opportunistic Infections, gaining weight.

Contrary to the data presented above the AIDS patients from the control group the following Symptoms: progressive decrease in total lympho cyte counts as well as in all T-lymphocyte subpopulations The number of T₄ cells increased the most advanced clinical pictures after about 10 months half off. There are five patients in this group meanwhile deceased and two patients who had an AZT Treatment had also shown one progressive decrease in T₄ lymphocytes. Thereby values  of 130 / mm³ and 250 / mm³ reached, one of the two patients has passed away. In this patient, one somewhat slower decrease in T₄ lymphocyte numbers on the Half watched. Two of the patients are in the WR-6 phase.

None of the seven HIV-1 infected patients orally RODILEMID® (30 to 18 months) has been treated has now reached a clinical picture. On the WR scale, they are on average between WR 1.66 and WR 1.33. Immunological examination has shown that in all T- Lymphocyte subpopulations increased in total were observed, with mean values of T₄ lymphocytes pay from 526 / mm³ to 825 / mm³ after four months of treatment could be observed.

AIDS kills children relatively soon. Of the four Children with the AIDS appearance who take orally Treating RODILEMID® for 26 months is one thing deceased, but the others are still alive.

In summary it can be stated here that RODILEMID® unambiguously to an improved clinical Appearance and extending the life of AIDS patients has contributed. In HIV-1 infected Patients were able to transition to the disease stage prevented or at least slowed down. The treatment is easy to do, it is inexpensive and it is have had no toxic or other side effects observed. The differences described above oral or intramuscular use with the non-toxicity of RODILEMID® allows the conclusion that RODILEMID® in double doses in adults can be administered, leading to even better ones Treatment success.

Claims (17)

1. Medicament containing

• (a) a polyaminopolycarboxylic acid or a derivative or a complex thereof;
• (b) an essential amino acid or a derivative thereof; and
• (c) a divalent element or a compound of a divalent element; or
• (a ′) a polyaminopolycarboxylic acid or a derivative or a complex thereof; and
• (b ′) a complex of an essential amino acid and a divalent metal ion;

with the exception of the combination

• (a) EDTA CaNa₂;
• (b) cysteine in a concentration of 0.38-2.3% by weight based on the sum of the three components;
• (c) calcium gluconate;

and optionally a pharmaceutically acceptable one Carrier. 2. Medicament according to claim 1, wherein the in (a) and (a ') called polyaminopolycarboxylic acid EDTA, DTPA, DCTA or a derivative or a complex or a salt thereof, is preferably EDTA-CaNa₂. 3. Medicament according to claim 1 or 2, wherein the in (b) and (b ′) amino acid called cysteine, histidine or Methionine and the derivative thereof is a precursor of Cysteine or histidine or of methionine or N- Acetylcysteine, N-acetylhistidine or N-acetylmethionine is.   4. Medicament according to one of claims 1 to 3, wherein the in (c) divalent element calcium and the Derivative thereof is a calcium salt, preferably calcium lactate, calcium gluconate or calcium levulinate. 5. Medicament according to one of claims 1 to 3, wherein the complex mentioned in (b ')

• (i) cysteine and copper;
• (ii) serine and copper;
• (iii) tyrosine and copper;
• (iv) cysteine and zinc;
• (v) serine and zinc; or
• (vi) tyrosine and zinc.

6. Medicaments containing at least one of the following mixtures

• 1. EDTA-CaNa₂, N-acetylcysteine and calcium gluconate;
• 2. EDTA-CaNa₂, calcium gluconate and cysteine in a concentration greater than 2.3 wt .-%, based on the sum of the three components;
• 3. EDTA-CaNa₂ and a complex of cysteine and copper;
• 4. EDTA-CaNa₂ and a complex of serine and copper;
• 5. EDTA-CaNa₂ and a complex of tyrosine and copper;
• 6. EDTA-CaNa₂ and a complex of cysteine and zinc;
• 7. EDTA-CaNa₂ and a complex of serine and zinc; or
• 8. EDTA-CaNa₂ and a complex of tyrosine and zinc.

7. Medicament according to one of claims 1 to 6, wherein the polyaminopolycarboxylic acid mentioned in (a) and (a ′) or the derivative or the complex thereof in one Concentration of 90-95 wt .-%, that mentioned in (b) essential amino acid or the derivative thereof in one Concentration of 0.3-3% by weight, the divalent element mentioned in (c) or the Connection of a divalent element in one Concentration of 3-7% by weight,  and the complex mentioned in (b ′) in a concentration of 3.3-7% by weight occurs. 8. Use of a medicament according to one of the claims 1 to 7 for the treatment of viral, preferably retro viral diseases. 9. Use of a medicament according to one of the claims 1 to 7, and a drug containing the in Disclaimer of claim 1 composition, for Treating AIDS. 10. Use of a medicament according to one of the claims 1 to 7, preferably from herpes simplex, herpes zoster and facial paresis. 11. Use of a medicament according to one of the claims 1 to 7 as an immunomodulator. 12. Use of a medicament according to one of the claims 1 to 7, and a drug containing the in Disclaimer of claim 1 composition for Treatment of autoimmune diseases. 13. Use according to claim 12, wherein the car immune disease multiple sclerosis or the guillaume Barre syndrome is. 14. Use of a medicament according to one of the claims 1 to 7 for the treatment of neurodegenerative Diseases. 15. Use of a medicament according to one of the claims 1 to 7 and a drug containing the in Disclaimer of claim 1 composition as anti-cancer drug.