Title:  Use of substance P antagonists in the treatment of the adenocarcinomas

United States Patent:  6,576,638

Issued:  June 10, 2003

Inventors:  Pompei; Pierluigi (Camerino, IT); Massi; Maurizio (Camerino, IT); Nabissi; Massimo (Tolentino, IT); Sparapani; Pier Luigi (Camerino, IT)

Assignee:  Biopolis S.p.A. (Siena, IT)

Appl. No.:  030305

Filed:  May 13, 2002

PCT Filed:  July 5, 2000

PCT NO:  PCT/EP00/06309

PCT PUB.NO.:  WO01/01922

PCT PUB. Date:  January 11, 2001

It is described the use of antagonists of neuro kynin receptors (NK-1; substance P receptors) in order to establish a new drug effective in the treatment of adenocarcinomas, such antagonists being one or more substances selected among those ones having the following features: pA₂ >6.0 both in human and in murine tissues, etherocyclic non peptidergic structures, antiangiogenic effects experimentally demonstrated onto the genito-urinary tract rumors induced via orthotopic drafts of human tumoral cells either in the genito-urinary apparatus of either immunodeficient rats or mice, decrease of the tumoral mass on tumors of the genito-urinary tract induced by orthotopic drafts of tumoral cells onto tissues of the genito-urinary tract of either immunodeficient rats or mice.

DETAILED DESCRIPTION OF THE INVENTION

The present invention refers to the use of substance P receptor antagonists particularly NK-1 receptor antagonists, in the treatment of adenocarcinomas, particularly genito-urinary tract neoplasms, more particularly prostatic carcinoma. Selected antagonists according to the present invention are listed in Tab. 1 and are characterised by the following features:

1) pA₂ >6.0 both in murine and human tissues, pA₂ being the concentration of the drug at which it is half-maximally effective, and the pA₂ is directly related to the affinity of the ligand to the receptor,

2) Non peptidic heterocyclic structure.

3) Antiangiogenic effects experimentally evaluated onto tumors of the genito-urinary tract induced via orthotopic grafts of tumoral human cells on tissues of the genito-urinary tract of rats and/or immunodeficient mice. Orthotopic graft is intended to be a graft of cells in the host, via direct injection of cells.

4) Reduction of the tumoral mass experimentally demonstrated onto tumors of the genito-urinary tract induced via orthotopic grafts of tumoral human cells on tissues of the genito-urinary tract of rats and/or immunodeficient mice.

The following substances are considered by the authors effective against adenocarcinomas, specifically against the adenocarcinomas originating from the genito-urinary tract.

 

The above mentioned substances may be used to prepare drugs in combination with known adjuvants. Sinergistically, they can be combined with substances listed in table 2. As a matter of fact, the combination of one or more substances listed in table 1 with one or more substances in table 2 (or corresponding derivatives of the substances listed in table 2, derivatives known to the man skilled in the art) provides a therapeutic positive response higher than 10%, as stated in points 3-4 of the pharmacological characteristics of the NK-1 antagonists.

 

Route of administration is based upon the specific characteristics of the compounds and implies the endovenous, intrabladder, intraperitoneal, intramuscular, subcutaneous and oral administration.

Adjuvants are selected among those commonly used in pharmacotherapy, such as: methyl-p-hydroxybenzoate, latex and saline solution.

Administration of substances listed in table 1, possibly in combination with substances listed in Table 2, is able to either reduce or reverse tumor growth via inhibition of angiogenesis and tumoral mass.

Moreover, use of antagonists of substance P is endowed of the following advantages:

Antiemetic effect, opposite to chemiotherapic drugs which show marked emetic effects

Antidepressive effect, which is a very important psychotherapic effect in cancer affected patients.

The following examples should be considered as illustrative of the present invention an not limitative of the scope of the invention itself.

MATERIALS AND METHODS

Animals

For this studies has been used male athymic nude mices (Harlan). The mices were housed in laminar flow cabinet under pathogenic free conditions and used at 4-5 weeks of age.

Orthotopic Implantation

The PC-3 human prostatic cancer cell lines (ECACC) were mantained in Minimum essential medium (GIBCO BRL) For in vivo studies, tumor cells in exponential growth phase were harvested by a 120 seconds treatment with tripsyn in 0.02% EDTA. After that the cells were resuspended in saline solution. 20 .mu.l (10⁵ cells) of saline solution was inoculated in nude mice prostate. The tumors were taken at different times (three up to 180 days) for analysis.

Treatments

Animals were divided in groups (10 animals each one) and treated with different solutions:

physiological solution, -Sustance P (1 ng up to 1 mg), -L-703,606 (1 ng up to 1 mg), -L-703,606+Leuprolide Acetate (using dosages reported in table 1 and 2).

The treatments were made at different time (one at day until one at week). For different periods of time (three days up to 180 days).

Immunohystochemistry

Frozen sections of tumors have been fixed in Acetone, acetone/chloroform, acetone.

The presence of new blood vessels has been highlighted using the Rat anti-Mouse CD31 antibody, a Goat anti-Rat POX as a secondary antibody and DAB as a chromogen substrate. For quantification of blood vessels an image analysis system was employed.

Cancerogenicity

The animals were maitained under observation for three up to 180 days. Autoptic examinations were performed in all animals and the tumoral mass weighed.

Results

Vascular Density Quantification

The quantitative data obtained on blood vessels density have shown a statistically significant difference (p<0.05) amongst different treatments. SP treated mice group showed a statistically significant increase of CD 31 values compared to control groups (p<0.05).

Moreover, a statistically significant reduction (p<0.05) of CD 31 values has been outlined in mice treated with L-703,606 compared either to control groups or SP treated mice. The inhibitory angiogenic effect mediated by L-703,606 has been shown in all groups treated with this compound. A statistically significant reduction of CD 31 values has been shown with L-703,606+Leuprolide, Acetate (p<0.05) compared to L-703,606 groups (>10%).

Tumoral Mass Weight

Tumoral mass weight has been evaluated in all experimental groups, throughout treatment days (from 3 up to 180 days). Results have shown a statistically significant reduction (p<0.05) of tumoral mass weight in mice treated with L-703,606, compared to either control groups or SP treated mice. Moreover, combination of L-703,606 with Leuprolide Acetate allows to reveal a further tumoral mass reduction, (>100%).

Correlation between tumoral mass weight and type of treatment is proportionally related to treatment time-course.

Claim 1 of 11 Claims

What is claimed is:

1. A method for treating adenocarcinomas comprising the steps of administering to a subject in need of treatment a pharmaceutically effective amount of an antagonists of NK-1 receptors, such antagonists being one or more compounds and corresponding mixtures selected among the compounds having the following characteristics:

pA₂ >6.0 in human and murine tissue;

non peptidic heterocyclic structure;

antiangiogenetic effects experimentally demonstrated in urogenital tumors, induced by orthotopic implantation of human tumoral cells in immunodeficient rat and mice urogenital tissues, said orthotopic implantation being made by inoculating the rat and mice urogenital tissues with said human tumoral cells and said angiogenetig effects being measured by the reduction of tumoral mass in the urogenital tumors induced by the orthotopic implantation.
 

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