University of Alicante

The Institute of Organic Synthesis of the University of Alicante has developed a procedure to synthesize a new family of molecules suitable for use in the prevention or treatment of cancer due to their cytotoxic properties.

The present invention refers to the zwitterion of saturated, monounsaturated, polyunsaturated and carboxylic 2-phosphocolinic acids and their pharmaceutically acceptable derivatives, with a structure described by the general formula (1):

• A = saturated, monounsaturated or polyunsaturated carbonate chain, with any configuration in the double bonds C=C, mono or polyhydroxylated in different positions, or non-hydroxylated.

• R = An atom or group of atoms weighing between 1 and 200 Dal, and their pharmaceutically stable derivatives (e.g. salts, hydrates or polymorphs).

The novel synthesis procedure for this family of compounds consists of two reaction steps (Schemes 1 and 2) under mild reaction conditions. The corresponding 2-hydroxy derived, obained from saturated, monounsaturated, polyunsaturated acid or its derivatives is transformed into the Zwitterion of saturated, monounsaturated or polyunsaturated 2-phosphocolinic carboxylic acid or one of its derivatives:

(i) Generation of an intermediate compound of general formula (2) by the addition of 2-bromoethyl dichlorophosphate of formula (3), or its chlorinated or iodinated analogue, on the hydroxyl group in alpha position to the carbonyl group, of a compound of formula (4) in the presence of a nonpolar solvent at ambient temperature.

Scheme 1. Generation of the intermediate compound of general formula (2). Both in formula (2) and in (4) A and R have the meanings given for the compound formula (1).

(ii) Replacement of the bromine atom (chlorine or iodine) of the intermediate compound of formula (2) by a trimethylamino group and formation of the corresponding betaine is performed. For this, the intermediate of the previous step (2) is reacted with trimethylamine dissolved in an alcohol, in mild conditions between 0ºC and 60ºC for 1-5 days and subsequent treatment with a mild inorganic base.

Scheme 2. Substitution reaction to form the corresponding betaine of the compound with general formula (1).

It has been observed that the general formula molecule (1) shows in vitro activity against different types of carcinogenic cells, for example against tumour cell lines: lung (non-small cell), pancreas, kidney, central nervous system (CNS), prostate, colon, breast, melanoma, ovary, acute lymphoblastic leukaemia, acute promyelocytic leukaemia, chronic myelogenous leukaemia, myeloma, large cell immunoblastic lymphoma, Burkitt lymphoma, non-Hodgkin's B-cell lymphoma, chronic lymphocytic leukaemia, mantle cell lymphoma, multiple myeloma and acute T-cell leukaemia.

The main advantages of this technology are the following:

• The synthesis of these compounds takes place under mild reaction conditions.

• The reaction conditions used improve the synthetic operation on a larger scale and the execution times are shorter.

• The products obtained do not present neither problems of solubility in water nor toxicity.

• The series of molecules obtained present preventive or curative activity for a great variety of carcinogenic cell lines (IC50 up to 7.06±1.45 µM).

• The synthesized zwitterions also present less degradation in the biological environment, which implies a greater therapeutic effect, as well as greater specificity towards the lipidic bi-layer of the cells and its action on them.

The main innovative aspect of this technology is the invention of a new family of zwitterionic molecules of saturated, monounsaturated, polyunsaturated acids-phosphocholine and their acid derivatives with the general structure (1) - whose characteristics confer the molecule a preventive or curative activity in the micromolar range for a wide variety of carcinogenic cell lines.

In addition, these new chemical formulations have been achieved using a simple and inexpensive synthetic route.

A series of compounds with the general formula (1) have been synthesised on a laboratory scale.

In a particularly preferential feature of the invention, A is a fragment of fifteen carbon atoms containing a double C=C bond between the C9-C10 carbons with Z configuration and the group R represents an ethyl ester COOEt (molecule MCH-811). Antitumour activity has been studied by measuring in vitro the IC50 index of different human tumour cell lines (pancreas, lymphoma, lung, kidney, etc.).

The current invention is framed as much in the field of the pharmaceutical chemistry as in the medicine area, with more specific emphasis to those compounds useful as cytotoxic agents.

Formula compounds (1) and their pharmaceutically acceptable derivatives such as salts, hydrates or polymorphs, can be used as active species against different types of carcinogenic cells, preferably in the prevention and/or treatment of cancers such as those mentioned in the technical description.

The research group is looking for companies interested in acquiring this technology for commercial exploitation through:

• Patent licensing agreements.

• Technical cooperation (R&D projects) to develop new molecules, new applications, carry out industrial scaling, adaptation to the specific needs of companies, etc.

• Etc.

This technology is protected by patent application.

• Title of the patent: "Zwitterionic compounds of carboxylic 2-phosphocolinic acids and their use as cytotoxic agents".

• Application number: P201830802

• Application date: 02/08/2018