COMBINATION COMPRISING A NEUROTENSIN RECEPTOR BINDING COMPOUND AND NAPOLI

Abstract

The present invention relates to a combination comprising a neurotensin receptor binding compound and NAPOLI for use for the treatment of a neurotensin receptor overexpressing tumour in a subject.

Claims

1. A method of treating a neurotensin receptor overexpressing tumor, comprising administering to a subject in need thereof a therapeutically effective amount of a combination comprising a neurotensin receptor binding compound, folinic acid, fluorouracil, and liposomal irinotecan (nal-IRI).

2. The method of claim 1, wherein the neurotensin receptor binding compound is radiolabeled with a therapeutic radionuclide.

3. The method of claim 2, wherein the therapeutic radionuclide is selected from the group consisting of .sup.177Lu, .sup.90Y .sup.67Cu, .sup.131I, .sup.186Re, .sup.188Re .sup.211At, .sup.212Pb, .sup.213Bi, .sup.225Ac, and .sup.227Th.

4. The method of claim 2, wherein the therapeutic radionuclide is .sup.177Lu or .sup.225Ac.

5. The method of claim 1, wherein the neurotensin receptor binding compound comprises a molecule of formula (i): ##STR00003## or a complex thereof.

6. The method of claim 1, wherein the neurotensin receptor binding compound is radiolabeled with .sup.225Ac.

7. The method of claim 1, wherein the neurotensin receptor binding compound is a complex of formula (ii): ##STR00004##

8. The method of claim 1, wherein the neurotensin receptor overexpressing tumour is pancreatic ductal adenocarcinoma or colorectal cancer.

9. The method of claim 1, wherein the neurotensin receptor binding compound is an NTR1 binding compound.

10. The method of claim 1, wherein the neurotensin receptor binding compound is administered in simultaneous, separate, or sequential combination with folinic acid, fluorouracil, and liposomal irinotecan.

11. The method of claim 2, wherein the radiolabeled neurotensin receptor binding compound is administered by injection IV at a dose of about 2 to 7 GBq per injection.

12. The method of claim 2, wherein the radiolabeled neurotensin receptor binding compound is administered as a unitary dosage of less than 40 MBq.

13. The method of claim 2, wherein the therapeutic radionuclide is a beta-emitting radionuclide or an alpha-particle emitting radionuclide.

14. The method of claim 1, wherein the neurotensin receptor overexpressing tumor is selected from the group consisting of pancreatic ductal adenocarcinoma (PDAC), small cell lung cancer, prostate cancer, colorectal cancer, breast cancer, meningioma, Ewing's sarcoma, pleural mesothelioma, head and neck cancer, non-small cell lung cancer, gastrointestinal stromal tumors, uterine leiomyoma, and cutaneous T-cell lymphoma.

15. The method of claim 1, wherein the subject has metastatic PDAC and has not previously received a therapy for treating pancreatic cancer.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0080] FIG. 1 represents the administration schedules from Groups 1 to 6.

[0081] FIG. 2 represents the tumor volume of mice bearing subcutaneous AsPC1 tumors from Groups 1, 2, 3 and 5 (Group 1=black dots; Group 2=squares; Group 3=triangles; Group 5=inverted triangles), wherein Group 5 was treated with a combination according to the invention.

[0082] FIG. 3 represents the tumor volume of mice bearing subcutaneous AsPC1 tumors from Groups 1, 2, 4 and 6 (Group 1=black dots; Group 2=squares; Group 4=pentagons; and Group 6=stars), wherein Group 6 was treated with a combination according to the invention.

EXAMPLES

Materials & Methods

Reference Substances

[0083] Leucovorin (folinic acid, Ca.sup.2+ complex) was dissolved with 0.9% NaCl solution to reach at 25 mg/mL final concentration. 5-FU stock solution was diluted with 0.9% NaCl solution to reach 25 mg/mL concentration. Liposomal irinotecan stock solution 5 mg/mL was diluted with 1?PBS to reach at 3.2 mg/mL concentration.

Radiolabeling of IPN01087 with Lutetium-177

[0084] The radiolabeling procedure was performed using ammonium acetate 0.4 M containing 0.325 M gentisic acid pH 4.2 buffer and lutetium-177 (.sup.177LuCl.sub.3, ITG, specific activity >3,000 GBq/mg at calibration).

[0085] In a microtube, lutetium-177 (.sup.177LuCl.sub.3) was mixed with ammonium acetate 0.4 M containing 0.325 M gentisic acid pH 4.2 buffer (2.8? volume of lutetium-177 solution) and IPN01087 to reach a specific activity of 85 MBq/?g. The reaction mixture was incubated at +85? C. for 30 minutes using a heating system. At the end of the incubation period, the radiolabeling incorporation was assessed by reversed phase liquid chromatography and thin layer chromatography. The radiolabeling mixture was then diluted with ammonium acetate 0.4 M containing 0.325 M gentisic acid pH 4.2 buffer, 0.9% NaCl solution, and DTPA Ca(Na.sub.3) to reach the desired radioactive doses in MBq and concentrations.

Vehicles

[0086] The vehicle of irinotecan is hereafter referred as vehicle #1. The vehicle of 5-FU and Folinic acid is hereafter referred as vehicle #2. The vehicle of .sup.177Lu-IPN01087 is hereafter referred as vehicle #3.

Example 1: Efficacy Study on Animals Induced with AsPC1 Cells

Cancer Cell Line

[0087] The AsPC1 cell line was isolated from a metastatic site (ascites) from a 62-year old female patient (Chen WH. et al., In Vitro. 1982 January; 18(1):24-34).

Cell Culture Method

[0088] Tumor cells were grown as monolayer at 37? C. in a humidified atmosphere (5% CO2, 95% air). The culture medium is RPMI 1640 containing glutamax supplemented with 10% heat inactivated fetal bovine serum, sodium pyruvate and hepes. For experimental use, tumor cells were detached from the culture flask using accutase and neutralized by addition of complete culture medium. The cells were counted and their viability was assessed by 0.25% trypan blue exclusion assay. Two frozen pellets of AsPC-1 tumor cells were prepared: one frozen cell pellet prepared during the in-vitro cell culture, and one frozen cell pellet prepared using the cell suspension used for tumor induction in mice.

Animals

[0089] One hundred fifty (150) healthy female SWISS Nude mice (Crl:NU(Ico)-Foxn1nu), 7 weeks old at reception, were obtained from Charles River.

Tumor Induction

[0090] Tumors were induced by subcutaneous injection of ten million (107) AsPC-1 cells in 200 ?L of RPMI 1640 medium containing matrigel (50:50, v:v, ref: 356237, BD Biosciences, France) into the right flank (in the axis of the heart) of 150 female animals. AsPC-1 tumor cell implantation was performed 72 hours after a whole body irradiation with a gamma-source (2 Gy (Nude mice), .sup.60Co, BioMep, France). The day of randomization was designated as the day 0 (DO). Animals were grafted with AsPC-1 tumors a week prior to randomization (D-7).

Randomization

[0091] Animals were randomized by individual tumor volume when tumors reached a mean volume of 150-200 mm.sup.3. Forty-eight animals (48) out of 150 were randomized into 6 groups of 8 animals each using Vivo Manager? software (Biosystemes, France). Homogeneity between groups was tested by an analysis of variance (ANOVA).

Treatment

[0092] .sup.177Lu-IPN01087 and the reference substances were administered by intravenous injection (IV) into the caudal vein via a catheter or by intraperitoneal (IP) injection. The recommended pH formulation for IV administration is pH 4.5-8.0 and for IP administration 7.3-7.4. The administration volumes were as follows: [0093] Group 1 (control group): 5 mL/kg for vehicle #1, 4 mL/kg for vehicle #2, 50-70 ?L/mouse for vehicle #3, and 50-70 ?L for flushing the catheter. [0094] Group 2 (NAPOLI group): 5 mL/kg for Irinotecan, 2 mL/kg for 5-FU, 2 mL/kg for Folinic acid, 50-70 ?L/mouse for vehicle #3, and 50-70 ?L for flushing the catheter [0095] Groups 3 and 4 (.sup.177Lu-IPN01087 groups): 5 mL/kg for vehicle #1, 4 mL/kg for vehicle #2 and 50-70 ?L for .sup.177Lu-IPN01087 and 50-70 ?L for flushing the catheter. [0096] Groups 5 and 6 (NAPOLI+.sup.177Lu-IPN01087 groups): 5 mL/kg for Irinotecan, 2 mL/kg for 5-FU, 2 mL/kg for Folinic acid, 50-70 ?L for .sup.177Lu-IPN01087, and 50-70 ?L for flushing the catheter.

[0097] The animal groups were treated as follows: [0098] Group 1 (control group): animals received one weekly IV injection of vehicle #1 for a total of 3 injections combined with one weekly IP injection of vehicle #2 for a total of 3 injections, and combined with one weekly IV injection of vehicle #3 for a total of 3 injections. [0099] Group 2 (NAPOLI group): animals received one weekly IV injection of liposomal irinotecan at 16 mg/kg for a total of 3 injections, combined with one weekly IP injection of Folinic acid at 90 mg/kg for a total of 3 injections plus one weekly IP injection of 5-FU at 50 mg/kg for a total of 3 injections, and combined with one weekly IV injection of vehicle #3 for a total of 3 injections. [0100] Group 3 (.sup.177Lu-IPN01087? 20MBq group): animals received once weekly IV injection of vehicle #1 for a total of 3 injections, combined with one weekly IP injection of vehicle #2 for a total of 3 injections, and combined with one weekly IV injection of .sup.177Lu-IPN01087 at 20 MBq/mouse (0.443 nmol/mouse) for a total of 3 injections. [0101] Group 4 (.sup.177Lu-IPN01087? 35MBq group): animals received once weekly IV injection of vehicle #1 for a total of 3 injections, combined with one weekly IP injection of vehicle #2 for a total of 3 injections, and combined with one weekly IV injection of .sup.177Lu-IPN01087 at 35 MBq/mouse (0.776 nmol/mouse) for a total of 3 injections. [0102] Group 5 (NAPOLI+.sup.177Lu-IPN01087? 20MBq group): animals received one weekly IV injection of liposomal irinotecan at 16 mg/kg for a total of 3 injections, combined with one weekly IP injection of Folinic acid at 90 mg/kg for a total of 3 injections plus one weekly IP injection of 5-FU at 50 mg/kg for a total of 3 injections, and combined with one weekly IV injection of .sup.177Lu-IPN01087 at 20 MBq/mouse (0.443 nmol/mouse) for a total of 3 injections. [0103] Group 6 (NAPOLI+.sup.177Lu-IPN01087? 35MBq group): animals received one weekly IV injection of liposomal irinotecan at 16 mg/kg for a total of 3 injections, combined with one weekly IP injection of Folinic acid at 90 mg/kg for a total of 3 injections plus one weekly IP injection of 5-FU at 50 mg/kg for a total of 3 injections, and combined with one weekly IV injection of .sup.177Lu-IPN01087 at 35 MBq/mouse (0.776 nmol/mouse) for a total of 3 injections.

[0104] 20MBq in mice is equivalent to about 4.7 GBq in human (human equivalent dose).

[0105] 35MBq in mice is equivalent to about 7.6 GBq in human (human equivalent dose).

[0106] At the time of tumor relapse (or at termination if no escape occurs), the tumor from 3 out of 8 mice from each group were collected. Tumors will be weighed, flash-frozen and then stored at ?80? C.

[0107] The treatment efficacy was assessed in terms of the effects of the treatments on the tumor volumes of treated animals relative to control animals. The tumor volume was estimated by the formula:

[00001]$\mathrm{Tumor}\mathrm{Volume}=\frac{{\mathrm{width}}^2?\mathrm{length}}{2}$

Tumors which were palpable and not measurable using calipers were assigned a volume of 4 mm.sup.3, indicating the technical limit measure. Tumor volume of 1000 mm.sup.3 were considered to be equal to 1 g. Individual, mean and median tumor volumes were measured.

[0108] Results are presented in FIGS. 2 and 3.

[0109] FIG. 2 shows AsPC-1 tumor growth in mice treated with vehicles (Group 1=circles), NAPOLI (Group 2=squares), .sup.177Lu-IPN01087 @20MBq (Group 3=triangles), or a combination of NAPOLI and .sup.177Lu-IPN01087 @20MBq (Group 5=inverted triangles). While NAPOLI treatment alone produced an antitumor effect when compared to the vehicles, the combination of NAPOLI and .sup.177Lu-IPN01087 significantly reduced tumor volumes as compared to either single treatment.

[0110] Furthermore, the data shown in FIG. 3 demonstrate that a combination of NAPOLI and .sup.177Lu-IPN01087 at a higher dose of 35MBq (Group 6=stars) displayed even better reduced tumor volumes control as compared to NAPOLI (Groupe 2=squares) and .sup.177Lu-IPN01087 (Group 4=pentagons) alone.

[0111] Overall, .sup.177Lu-IPN01087 radiotherapy in combination with the NAPOLI chemotherapy regimen showed improved anti-tumoral efficacy in AsPC-1 pancreatic cancer models as compared to the NAPOLI chemotherapy or radiotherapy regimens alone.