DE3245188C2 - 1-N- [2- (2-dimethylaminoethoxy) acetylamino] adamantane phosphonoacetate, its production process and pharmaceutical compositions containing this compound - Google Patents

Abstract

1-N- [2- (2-dimethylaminoethoxy) acetyl-amino] adamantane phosphonoacetate, the structure of which is described below, has a synergic anti-viral effect as compared with the effect of one component. (1st formula).

Description

The present invention relates to a new compound with anti-viral action, namely 1-N- [2- (2-dimethylaminoethoxy) acetyl-amino] -adamantane-phosphonoacetate, which has the following structural formula »:

NH-CO-CHj-O-CH ₂ -CH ₂ -N (CHj) ₂ "

HOOC - CHj - POjHj (I)

The anti-viral effect of phosphonoacetic acids against vaccinia viruses, herpes type 1 and herpes type 2 is known for a long time. In this context, reference is made to the following literature: N. L Shipkowitz et al, Appl. Microb. 26: 264-267 (1973); J. E. Sugar et al. Invest. Ophthalmol. 1973, 12: 532-534; D. D. Gerstein et al, Antimicrob. Agents and Chemoth, 7,285-288 (1975); R. F. Meyer, ibid. 9,308-311 (1976).

The effect of l-N- [2- (2-dimethylaminoethoxy) acetylarninoJadarnantan - also called tromantadine - against the same viruses is also described (D. Peteri & W. Sterner, Arzneimittelforsch. 23, 577 (1973); G. May & D. Peteri, ibid. 23,718 (1973)).

It has now been found that compound (I), or tromantadine phosphonacetate, has a surprisingly high Has activity against herpes viruses compared to the above two components as well as one Effect in the same order of magnitude against vaccinia viruses. The present invention describes and includes also pharmaceutical agents with anti-viral effects which contain compound (I) as an active ingredient.

The invention also relates to the manufacturing process of tromantadine phosphonoacetate, thereby characterized in that phosphonoacetic acid and tromantadine in a molar ratio of substantially 1: 2 in polar solvents such as water, or preferably lower alcohols, to react with one another be brought and that the salt thus formed by evaporating the solution or by adding Precipitants is isolated. The reaction is carried out at room temperature. The following example illustrates this method according to the invention.

example

a) 1-N- (2-chloracciyl-amino) adamantane
50 g of 1-aminoadamantane hydrochloride are in a 10% aqueous solution of 21.3 g of sodium hydrochloride

xid given; then, while stirring using a magnetic stirrer and cooling with ice to below 5 ° C., a solution of 213 ml of Chk> racetyl chloride in 100 ml of methylene chloride are added. The reaction mixture is left as it is for 2 hours with circular stirring; the organic phase is separated off, washed with water and washed with Magnesium sulfate dried, the solvent is evaporated off under reduced pressure. The residue is recrystallized with diisopropyl ether: isopropanol 80:20. The desired product is obtained with a yield of 80%, the melting point is 122-123 ° C.

b) 1 -N- [2- (2-dimethylaminoethoxy) acetyl-amino] adamantane (tromantadine)

To a solution of 313 g (035 mol) of dimethylaminoethanol in 150 ml of anhydrous tetrahydrofuran ίο 8.4 g (035 mol) of sodium hydride are added while stirring and maintaining a temperature of 15-20 ° C When no more hydrogen bubbles rise, 40 g (0.17 mol) of 1-N- (2-chloroacetylamino) adamantane are obtained added, the whole is refluxed for 15 hours and then poured onto ice. The organic Phase is extracted with ethyl ether, washed with water and dried with potassium carbonate. The solvent is evaporated off under reduced pressure and the residue is chromatographed over silica gel purified, eluting first with diethyl ether then with ethanol. The pure product (dense yellowish oil) is isolated with a yield of 53%.

c) tromantadine phosphonoacetate (I)

20 g (0.142 mol) of phosphonoacetic acid and 80 g (0.285 mol) of tromantadine are dissolved in 120 ml of absolute ethanol dissolved The solution obtained is evaporated to dryness under vacuum. The result is a low-melting crystalline solid (melting point 33-40 ° C), which is very hygroscopic and in water, methanol and Ethanol is very soluble.

Elemental analysis for CmH ₆ IN ₄ PO ₉ (molecular weight = 700.83)

Calculated% C 58.26; H 8.77; N 7.99 Found% C 57.78; H 9.04; N 7.82

IR spectrum (in Nujol; the details of the absorption bands are in cm - '):

Binding broad bands between 3550 and 2500 OH

Bonds C = O (acid)

broad band between 1700-1670 C = O (amide)

NMR H ¹ spectrum (recorded in CDCI3, internal standard TMS; the chemical shift values are expressed in J):

2.4 (s, 12H, 2 (CH3) 2); From 2.6 to 3 (m, 4 H, 2 (CH ₂ -N));

33-3.9 (m, 10H, 2 (CO-CH ₂ -O); 2 (CH ₂ -O);

P-CH ₂ - COOH);

6.8 (s, 1 H, mobile COH);

8.3 (s, 4 H, mobile NH).

The toxic-pharmacological properties of tromantadine phosphonacetate (here for the sake of brevity AFP 723) are summarized as follows:

Celloxicity

The neutral red incorporation method was used, which provides an indication of the viability of cells in contact with the drug.

In order to obtain a more precise determination of the type and extent of the inhibition, the following were also made Valued parameters:

a) growth time and cell division capacity;

b) incorporation of ³ H thymidine in the DNA;

c) incorporation of ³ uridine into the RNA;

d) Storage of ¹⁴ C leucine in proteins.

The results obtained have shown toxicity up to values of 250 μg of the drug / ml of medium. Above the concentrations, there is a gradual decrease in terms of the storage of the used

th radioactive indicator with an approximately 50% inhibition of protein synthesis over 500 μg / ml.

The inhibition of the DNA appears to be less strong; this increases at concentrations above 100 μg / ml 50% reduced

Human diploids (WI-38) were found in primary or secondary cultures and monkey kidney cells in used continuous cultures

The toxicity values given above are given in view of the high pharmacological activity that will be described later under the section named with anti-viral effects, in a very favorable therapeutic Experience implemented

ίο Local tolerances

These were determined by using an aqueous solution containing 5% of the compound and a 5% ointment in white petroleum jelly.

Both the aqueous solution and the ointment were placed on the back twice a day for 4 weeks New Zealand rabbits shaved with an electric razor applied where possible

Erythema and edema were examined daily; only appeared on the 20th day of treatment with a minor one The percentage (10%) of the rabbits treated with the aqueous solution was a slight temporary one Erythema, which did not increase in the following days until the end of the experiment.

absence of systemic effects

The same rabbits as in the previous experiment were used at the end of the 4 week treatment possible effects on circulation and respiration were investigated. In addition, hematochemical and main biochemical parameters determined. The main organs were removed and subjected to a histological examination Subject to examination. No effect on circulation and respiration was found here; the hematochemical and biochemical parameters were within normal limits, which histological examination revealed no pathological change.

Anti-viral activity

Human diploid cells (WI 38) cultured in Petri dishes and monkey kidney cells were used. Three DNA-containing viruses were used as infecting viruses (Vaccinia, Herpes I and Herpes 2) and three RN Α -containing viruses (influenza virus A, poliovirus, sync. uvirus).

The AFP 723 compound of the invention inhibits DNA-containing vaccinia and herpes viruses when concentrated from 1-5 µg / ml and 100 µg / ml or with a TCID50 at approximately 50 µg / ml.

If the cells are pretreated with 5 μg / ml of AFP 723 for 15 hours, the decrease in infectivity is 99.9% (from 10 ⁶ to <10 ³ ).

The anti-viral effect of the compound is exerted intracellularly, during an intermediate state viral morphogenesis, as demonstrated by examination with the electron microscope. At concentrations of 50-100 μg / ml, AFP 723 blocks the vaccinia virus if it is not already completely and Herpes 1 and Herpes 2 viruses during the intranuclear capsular state. A concentration from 10-50 μ ^ / πιΙ completely inhibits the production of stain-forming viruses.

The following overview shows the above data in a diagram, in comparison with equimolecular doses of phosphonoacetic acid (PPA) and tromantadine.
45

Infecting virus Vaccinia Herpes I Herpes 2

PPA + + + ++ + +

Tromantadir. + + + + + + + + +

The anti-viral activity of AFP 723 on herpes viruses is surprisingly high compared to that of the Drugs that were to be used for comparison, while activity on vaccinia virus in the same Order of magnitude.

These characteristics suggest that AFP 723 is useful in the treatment of herpes viral infections and especially for herpes labialis, herpes genitalis, herpes cornealis and herpes zoster would have to.

The present invention also relates to all industrially applicable aspects connected with the

bo use of AFP 723 for treatment of herpes virus infections. Therefore there is an essential Aspect of this invention in pharmaceutical formulations containing predetermined amounts of AFP 723.

This compound can be applied topically, e.g. B. in the form of a cream, a gel, a pen or a solution, with a concentration between 0.5 and 5% of the active element.

The vaccinia virus used in the tests was chickenpox virus.

Claims (4)

Patent claims: 1. The compound l-N-p - ^ - dirnethylarninoethoxyJacetyl-aminoJ-adamantan-phosphonacetat, which the has the following structural formula: NH-CO-CH ₂ -O-CH ₂ -CHj-N (CHj) ₁ ' HOOC-CH ₂ -PO ₃ H ₂ 2. Process for the preparation of the compound according to claim 1, characterized in that phosphonoacetic acid and l-N- [2- (2-dimethylaminoethoxy) acetyl-amino] adamantane (tromantadine) in one molar Ratio, which is substantially equal to 1: 2, with each other in polar solvents for reaction be brought, and that the salt thus formed by evaporating the solution or by adding Precipitants is isolated for this purpose. 3. The method according to claim 2, characterized in that the polar solvent is low molecular weight Alcohols are used. 4. Pharmaceutical agents for the treatment of herpes infections, especially herpes labialis. Herpes genitalis. Corneal herpes and herpes zoster which contain the compound claimed in claim 1 as active Contain active ingredient.