Aplidine treatment of cancers

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5 day, iv inf Optimal activities were observed in xenografted gastric (17-20%) and prostate (25-38%) tumours following ip administration. Follow-up studies necessitated using rats for iv infusions. With this variation, a 24 hr iv infusion schedule produced similar activities against HF gastric (20%) and HF prostate (31%) tumour cells. Cytotoxicity was also found using a 5 day iv infusion schedule against HF prostate tumour cells (33%). The extended in vivo evaluations not only show that there is a strong relative correlation to the in vitro cytotoxic profile, but also a strong correlation with in vivo models used to characterise the tumour selectivity that identified aplidine as a candidate for clinical development.

Example 3 A phase I and pharmacokinetic study of aplidine given as a weekly 24 hours infusion in patients with advanced solid tumours In vivo studies revealed that in vivo activity increased by prolonging infusion duration. In this study 16 patients were treated. patients characteristics: median age 55 years, median PS 1, male/ female 11 / 5, tumour types being as follows: Head and neck 5, kidney 2, colon 3, rectum 2, sarcoma 1 and melanoma 3, all pre-treated with chemotherapy (median 2 lines).

Aplidine was administered as a 24 h infusion at the following dose levels (Dls): 133 (3 pts), 266 (3 pts), 532 (3 pts), 1000 (3 pts), 2000 (3 pts) and 3000 (1 pt) mcg/m2/wk x 3 every 28 days.

No dose limiting toxicities (DLTs) were observed. Only mild non-haematological toxicities consisting of nausea g 1, mucositis g 1, asthenia g 1 were reported. Phlebitis of the infusion arm was common and concentration-dependent. Pharmacokinetic analysis was performed in all patients, showing plasma levels at the DLs 1000 mcg/m2/w and 2000 mcg/m2/w equivalent to the active in vitro concentration (1 ng/ml). At DL 532 mcg/m2/w clinical improvement was observed in 1 pre-treated patient with advanced melanoma.

Example 4 Phase I and pharmacokinetic (PK) study of aplidine (APL) using a 24 hour, weekly schedule.

To date, 25 pts (median age 58 yrs, median ECOG 1) with advanced, previously treated solid tumours or lymphoma have been treated in this phase I study. Using a schedule of APL 24 h weekly x 3 followed by 1 wk rest, the following dose levels have been tested: 133 (n-3 pts), 266 (3), 532 (3), 1000 (3), 2000 (3), 4500 (3) and 3750 ~g/m2/wk (3). With 60 cycles ( 180 infusions) administered, all pts are evaluable for toxicity.

The Maximum Tolerated Dose was 4500 ~g/m2/wk x 3 with grade (G) 3 muscular toxicity (biopsy proven type II muscular atrophy). G4 CPK

and G3 transaminitis the dose-limiting toxicities in 2 or 4 pts. G2 malaise was observed in most pts and G2/3 emesis at >_ 2000 ~.g/m2 phlebitis is common but concentration-dependent. All pts have been sampled for PK analysis by LC/MS/MS. Preliminary data indicate extensive tissue distribution, a long elimination t 1/2 of 10-24 h nad plasma levels > 1 ng/ml (which is active in vitro). One pt with advanced melanoma resistant to DTIC/interferon had a clinical improvement maintained for > 30 weeks. This weekly infusional study demonstrates the feasibility and activity of a dose-dense APL schedule. As expected, neuromuscular toxicity is dose-limiting. The possible recommended dose of 3750 ~g/m2/wk x 3 is being assessed.

Example 5 Clinical pharmacokinetics (PK) of aplidine (APL) in patients with solid tumours and non-Hodgkin lymphomas.

Four intravenous schedules are under phase I evaluation: weekly 24 h infusion, biweekly 24 h infusion and 5 consecutive days 1 h infusion every 3 weeks. APL blood concentrations are analysed by liquid chromatography-tandem mass spectrometry. Initial results show accumulation in blood cells and a plasma PK characterised by na extensive distribution (volume of distribution usually in excess of 200 L/m2) and elimination half lives in the order of 10 to 24 h. An open 2-compartment model fits appropriately most profiles after 24 h infusion.

For 1 h infusions a 3-compartment model provides a better fit in most cases. Obtained plasma levels are known to be active in vitro.

Evaluation of additional patients, and a comparison of blood cells and plasma PK is ongoing.

Example 6 Preliminary results of a phase I and pharmacokinetic study of aplidine given as a 24 hour infusion every 2 weeks in patients with solid tumours and non-Hodgkins lymphomas.

Aplidine is given as a 24 hour infusion every 2 weeks. The starting dose was 200 ~g/m2/d and dose escalation proceed including so far 400, 800, 1600, and 3200 ~g/m2/d. A total of 18 patients (M/F: 7/ 11, median age 52, OMS 0/ 1: 10/8) have been entered. So far, no dose limiting toxicity was observed. Among evaluable patients, toxicity consisted of mild grade I-II nausea/vomiting, grade I-II asthenia, and cramping occurring during or immediately after the infusion. No neuromuscular toxicity were reported at the evaluated doses. One patient with an advanced lung cancer and documented tumour progression at the dose of 1600 ~g/m2 developed a haemolytic anaemia and thrombocytopenia that was considered unlikely related to the study drug. Initial pharmacokinetic analysis showed that the drug is extensively distributed and blood concentrations. An open 2-comarptment model fits appropriately most plasma concentration profiles. The terminal half-life is usually in the order of 10-24 h.

Clinical improvement was seen in a patient with a non-Hodgl~in lymphoma. The accrual is ongoing to determine the dose limiting toxicity and the dose to be recommended in phase II studies.