Novel applications of 1,3-propanodiole derivatives

The subject of the invention is the use of a compound of formula I,

CH₂OR₃

And₅r <n-e-ch₂or₃CH — R, in which

R1 is an optionally substituted straight or branched carbon chain having from 12 to 22 carbon atoms, which can be optionally separated by optionally substituted phenylene, and each of R₂, R3, R4 and R5 are independently H, in free form or in the form of a pharmaceutically acceptable salt, for the preparation of a pharmaceutical composition for the prevention or treatment of graft vascular diseases in a recipient of an alo or xenograft organ or tissue.

Another object of the invention is the use of a compound of formula I,

CH₂OR₃

AND

R5R4N-e-ch₂or₃CH-R, in which

R1 is an optionally substituted straight or branched carbon chain having from 12 to 22 carbon atoms that can be optionally separated by optionally substituted phenylene, and R2, R3, R4 and R5 each independently is H, in free form or in pharmaceutically acceptable salt form , for the preparation of a pharmaceutical composition for preventing or controlling acute rejection in a recipient of a transplanted xenograft.

Preferably, the compound of formula I is 2-amino-2- [2- (4-octylphenyl) ethyl] -1,3-propanediol in free form or in the form of a pharmaceutically acceptable salt.

In another preferred embodiment, the compound of formula I is 2-amino-2- [2- (4-octylphenyl) ethyl] -1,3-propanediol hydrochloride.

Preferably, the pharmaceutical composition is used together with a pharmaceutical composition comprising an immunosuppressive or immunomodulatory drug.

Preferably, the immunosuppressant or immunomodulatory drug is selected from the group consisting of cyclosporin A, FK-506, azathioprine; methotrexate; mycophenolic acid; mycophenolate mofetil, rapamycin 40-O- (2-hydroxyethyl) -rapamycin, 15-deoxyspergualine; monoclonal antibodies to leukocyte receptors or their ligands, and CTLA4-Ig.

Preferably, the monoclonal antibodies to leukocyte receptors are selected from the group consisting of MHC, CD2, CD3, CD4, CD7, CD25, CD28, B7, CD40, CD45 or CD58 antibodies, or ligands thereof.

Combined use includes simultaneous and sequential administration.

The invention can be used for

1.1. Prevention or treatment of the symptoms of chronic rejection, e.g. to avoid, reduce or reduce chronic rejection in an recipient of an organ or tissue of an alo- or hetero-transplant, e.g. heart, lung, heart-lung transplant, liver, kidney or pancreas where the recipient is administered is a therapeutically effective amount of the compound of formula I in free form or in pharmaceutically acceptable salt form;

1.2. Prevention or treatment of transplant vascular diseases, e.g. transplant vasculopathy, atherosclerosis or atherosclerosis in a recipient of an organ or tissue of an alo- or hetero-transplant, e.g. heart, lung, heart-lung transplant, liver, kidney or pancreas, where recipients administering a therapeutically effective amount of the compound of formula I in free form or in the form of a pharmaceutically acceptable salt;

190 348

In a series of further specific or alternative embodiments, the present invention may also be used to:

2. Prevention or control of acute rejection in a recipient of a transplanted xenograft, e.g., a patient receiving a heart, lung, heart-lung xenograft together, kidney, liver, bone marrow, pancreatic duct, skin or cornea, comprising the step of administering to said recipient a therapeutically effective amount of a compound of formula I in free form or in the form of a pharmaceutically acceptable salt.

The usefulness of compounds of formula I in free form or in the form of a pharmaceutically acceptable salt in chronic rejection, as well as usefulness in the treatment of the diseases and conditions listed above can be demonstrated in animal tests, for example according to the methods described herein below as well as clinically, for example. when the transplanted organ or tissue may undergo regular biopsy control, and in the case of heart transplantation, additional ultrasound scanning.

A. Prevention of graft vascular disease

Experimental animals:

Inbred DA rats (RTl^and, donors) and Lewis (RT1¹, allograft recipients) weighing between 200 and 300 grams. Animals were given unrestricted access to food and water before and after surgery.

Carotid artery transplantation:

Rats were anesthetized with isofluorate (Abbott) (4-5% for initiation, 1.5-2% for maintenance) and after anesthesia, 300 pg of atropine sulfate was injected subcutaneously. The left artery was dissected. The artery was clamped up and down, and about 7-10 mm was removed. The fissure was bridged with an allograft in which cold anemia was also induced for 45 min. Ethilon 10/0 seams were used. Finally, the skin was joined with 4/0 stitches. If necessary, then the Alzet osmotic minipump (Alza Corp. Palo Alto, Calif.) Was implanted subcutaneously on the back (alternatively the animals were treated orally).

The rats were subjected to one of the following procedures: the compound of formula I alone at doses of 0.1 to 10 mg / kg or in combination with cyclosporine A at doses of 0.03, 0.3 or 1 mg kg '* day ¹administered for 8 weeks using either the Alzet subcutaneous implanted osmotic minipump or alternatively by oral administration. After 8 weeks, the rats were sacrificed, the carotid arteries perfused for 1 min. 0.1 M saline solution with phosphate buffer (PBS, pH 7.4), followed by 15 min. 2.5% glutaraldehyde in phosphate buffer (pH 7.4). Carotid arteries were then excised and stained in Giems solution for histological evaluation.

Morphometric analysis involves measuring the thickness of the muscle and intima. Qualitative analysis of morphological changes includes scoring on a scale of 0-3 adventitial infiltration of mono-nuclear cells and necrosis (Aquatic Warbler degeneration, cell hypertrophy), smooth muscle cell nucleus (SMC) numbers in the environment (0-10, <100,> 100 and »100 nuclei for scoring, 0, 1, 2 and 3, respectively), SMC necrosis (Aquatic Warbler degeneration and SMC hypertrophy) and infiltration of the intima by mononuclear cells (13).

In both experiments, the compounds of formula I, especially Compound A in the form of the hydrochloride, significantly inhibit graft infiltration and the formation of a new intima.

B. In vivo cardiac xenograft (hamster-to-rat) The rat-hamster-xenograft combination is a so-called incompatible combination. Rats do not have sufficient natural anti-hamster antibodies in sufficient quantity to obtain the immediate hyper-rejection that is observed in compatible combinations; however, rejection in untreated recipients occurs within 3-4 days by combination with complementary antibodies. This is histologically illustrated by the destruction of blood vessels, the exudation and extravasation of erythrocytes, and the influx of polymorphonuclear granulocytes; there are often signs of hemorrhage and thrombosis. If these rejections are overcome by effective inhibition of antibody synthesis or complementary inactivation, cellular rejection may emerge later. This is visualized histologically by the inflow of mononuclear cells, including lymphocytes, lymphoblastic cells, macro phages, and destruction of muscle fiber parenchyma, inhibition of the cell rejection process requires more immunosuppression than for allografts. Rats with congenital emphysema (mu / mu)

190 348 do not have a competing (thymus-dependent) cellular immune system and are generally unable to reject allograft. Such animals reject hamster xenografts within 3-4 days in a similar manner to eutymic rats, indicating that (at least in part) the synthesis of anti-hamster antibodies in rats occurs following a thymic-dependent B cell reaction. Such recipients are useful in hamster xenografts to assess rejection via thymus-dependent antibody-mediated rejection.

The Syrian hamster heart was transplanted heterotopically into the abdomen of a male rat lewis (RT1 ') with anastomosis between the donor and recipient aorta and the right pulmonary artery donor with the lower vena cava of the recipient. The transplant was monitored daily by palpation of the abdomen. Rejection was proposed in the event of disappearance of heartbeat. The animals were weighed weekly. In a given series of experiments, 28 days were taken as the end point. The animals were dissected; outside the transplant, weight was assessed and thymus, spleen, liver and vas deferens were examined histologically. Blood was collected and processed into serum for the determination of anti-hamster cytolytic erythrocytic antibodies and complementary haemolytic activity.

The compounds were dissolved in water and administered orally daily in a volume of 2 ml / kg body weight. Administration of 5-30 mg / kg / day of a compound of Formula I, e.g. Compound A in the form of the hydrochloride, led to prolonged graft duration in both atimic and eutymic recipients.

The daily doses required for the practical application of the method of the present invention vary depending on, for example, the compound of formula I employed, the host, the mode of administration, the severity of the condition being treated and, optionally, an immunosuppressive drug, which is used in parallel, e.g. CysA. A preferred daily dose is in the range of about 0.03 to 2.5 mg / kg per day, especially 0.1 to 2.5 mg / kg per day, e.g. 0.5 to 2.5 mg / kg per day, in a dose of single or divided doses. Appropriate daily dosages for patients are in the order of 1 to 100 mg (orally). Suitable unit dosage forms for oral administration contain from about 1 to 50 mg, usually 5 to 30 mg, of the active ingredient, e.g., Compound A, e.g. in the form of the hydrochloride salt, together with one or more pharmaceutically acceptable diluents or carriers. Alternatively, the compound of formula I in free form or in pharmaceutically acceptable salt form may also be administered two or three times a week, e.g. at a dose as indicated above.

The compounds of formula I may be administered by any conventional route, in particular enterally, e.g. orally, e.g. in the form of drinking solutions, tablets or capsules, or parenterally, e.g. in the form of infectious solutions or suspensions. Pharmaceutical compositions containing compounds of formula I may be prepared in a conventional manner, e.g. as disclosed in EP-A 1-627 406.

The compounds of formula I may be administered as a single active ingredient or in combination with other drugs during immunomodulating therapy or with other anti-inflammatory agents. For example, compounds of formula I may be used in combination with cyclosporins, rapamycins or askomycins, or their immunosuppressive analogues, e.g. cyclosporin A, cyclosporin G, FK-506, rapamycin, 40-0- (2-hydroxy) ethyl: rapamycin, etc .; corticosteroids; cyclophosphamide; azathioprine; methotrexate; breąuinarem; leflunomide; mizorbina; mycophenolic acid; mycophenol mofetil; 15-deoxyspergualine; monoclonal immunosuppressive antibodies, e.g., leukocyte receptor monoclonal antibodies, e.g. MHC, CD2, CD3, CD4, Cd7, CD25, CD28, B7, CD40, cD45 or CD58 or their ligands; or other immunomodulatory compounds, e.g. CTLA4-Ig.

If the compounds of formula I are administered in combination with another immunosuppressive / immunomodulatory therapy, e.g. to prevent or treat chronic rejection, as detailed above, the doses of the co-administered immunosuppressive or immunomodulatory compound will of course vary depending on the type of co-administered drug, e.g. whether it is it's a steroid or cyclosporin, from the particular drug used, from the condition being treated, and so on. Accordingly, the present invention provides in a further further aspect:

190 348

Preparation example: soft capsules. A compound of formula I, e.g., Compound A 30 mg

Polyethylene glycol 300 300 mg

Polysorbate 80_20 mg

350 mg total

Compounds of formula I in free form or in the form of a pharmaceutically acceptable salt are well tolerated at the doses required for use in accordance with the present invention. For example, the acute LD50 is> 10 mg / kg in rats and monkeys.

Department of Publications of the Republic of Poland. Circulation of 50 copies Price PLN 2.00