ANTITUMOR COMBINATIONS CONTAINING ANTI-CEACAM5 ANTIBODY CONJUGATES AND FOLFOX

Abstract

The present invention concerns antibody-conjugates comprising an anti-CEACAM5-antibody for use for treating cancer in combination with folinic acid, 5-fluoro-uracil and oxaliplatin (FOLFOX). The invention further relates to pharmaceutical compositions and kit-of-parts comprising an anti-CEACAM5-antibody in combination with folinic acid, 5-fluoro-uracil and oxaliplatin (FOLFOX) for use for treating cancer.

Claims

1. An immunoconjugate comprising an anti-CEACAM5-antibody for use for treating cancer in combination with folinic acid, 5-fluoro-uracil and oxaliplatin (FOLFOX).

2. The immunoconjugate for the use of claim 1, wherein the anti-CEACAM5-antibody comprises a CDR-H1 consisting of SEQ ID NO: 1, CDR-H2 consisting of SEQ ID NO: 2, CDR-H3 consisting of SEQ ID NO: 3, CDR-L1 consisting of SEQ ID NO: 4, CDR-L2 consisting of amino acid sequence NTR, and CDR-L3 consisting of SEQ ID NO: 5.

3. The immunoconjugate for the use of claim 1 or 2, wherein the anti-CEACAM5-antibody comprises a variable domain of a heavy chain (VH) consisting of SEQ ID NO: 6 and a variable domain of a light chain (VL) consisting of SEQ ID NO: 7.

4. The immunoconjugate for the use of any of claims 1 to 3, wherein the anti-CEACAM5-antibody comprises a heavy chain (VH) consisting of SEQ ID NO: 8 and a light chain (VL) consisting of SEQ ID NO: 9.

5. The immunoconjugate for the use of any of claims 1 to 4, wherein the immunoconjugate comprises at least one cytostatic agent.

6. The immunoconjugate for the use of claim 5, wherein the cytostatic agent is selected from the group consisting of radioisotopes, protein toxins, small molecule toxins, and combinations thereof.

7. The immunoconjugate for the use of claim 6, wherein the small molecule toxins are selected from antimetabolites, DNA-alkylating agents, DNA-cross-linking agents, DNA-intercalating agents, anti-microtubule agents, topoisomerase inhibitors, and combinations thereof.

8. The immunoconjugate for the use of claim 7, wherein the anti-microtubule agent is selected from the group consisting of taxanes, vinca alkaloids, maytansinoids, colchicine, podophyllotoxin, gruseofulvin, and combinations thereof.

9. The immunoconjugate for the use of claim 8, wherein the maytansinoids are selected from the group consisting of N2′-deacetyl-N2′-(3-mercapto-1-oxopropyl)-maytansine (DM1) or N2′-deacetyl-N-2′(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4), and combinations thereof.

10. The immunoconjugate for the use of any of claims 1 to 9, wherein the anti-CEACAM5-antibody is covalently attached via a cleavable or non-cleavable linker to the at least one cytotoxic agent.

11. The immunoconjugate for the use of claim 10, wherein said linker is selected from the group consisting of N-succinimidyl pyridyldithiobutyrate (SPDB), 4-(pyridin-2-yldisulfanyl)-2-sulfo-butyric acid (sulfo-SPDB), and succinimidyl(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC).

12. The immunoconjugate for the use of any of claims 1 to 11, comprising an CEACAM5-antibody, which comprises a heavy chain (VH) consisting of SEQ ID NO: 8 and a light chain (VL) consisting of SEQ ID NO: 9 (huMAb2-3), and which is covalently linked to N2′-deacetyl-N-2′(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4) via N-succinimidyl pyridyldithiobutyrate (SPDB).

13. The immunoconjugate for the use of any of claims 1 to 12, wherein the immunoconjugate is characterised by a drug-to-antibody ratio (DAR) ranging from 1 to 10.

14. The immunoconjugate for the use of any of claims 1 to 13, wherein the cancer is selected from the group consisting of colorectal, stomach, pancreas, and oesophagus cancer.

15. The immunoconjugate for the use of any of claims 1 to 14, wherein the immunoconjugate and FOLFOX are administered simultaneously to a subject in need thereof.

16. The immunoconjugate for the use of claim 15, wherein the immunoconjugate and FOLFOX are formulated (i) in a single pharmaceutical composition comprising the immunoconjugate and FOLFOX, or (ii) in the form of at least two separate pharmaceutical compositions, wherein at least one pharmaceutical composition comprises the immunoconjugate, and one or more pharmaceutical compositions comprise folinic acid, 5-fluoro-uracil and oxaliplatin, in separate or combined formulations.

17. The immunoconjugate for the use of any of claims 1 to 14, wherein the immunoconjugate and FOLFOX are administered separately or sequentially to a subject in need thereof.

18. The immunoconjugate for the use of claim 17, wherein the immunoconjugate and FOLFOX are formulated in the form of at least two separate pharmaceutical compositions, wherein (i) at least one pharmaceutical composition comprises the immunoconjugate, and (ii) one or more pharmaceutical compositions comprise folinic acid, 5-fluoro-uracil and oxaliplatin, in separate or combined formulations.

19. The immunoconjugate for the use of any of claims 1 to 18, wherein the immunoconjugate comprising an anti-CEACAM5-antibody, and folinic acid, 5-fluoro-uracil and oxaliplatin (FOLFOX) are administered in 8 to 16 cycles, wherein one cycle comprises: administering the immunoconjugate at a dose of from 60 to 210 mg/m.sup.2, at least once in the cycle; administering folinic acid at a dose of from 100 to 300 mg/m.sup.2 or L-folinic acid at a dose of 100 to 200 m/m.sup.2, at least once in the cycle; administering 5-fluoro-uracil at a dose of from 1000 to 2000 mg/m.sup.2, at least once in the cycle, and administering oxaliplatin at a dose of from 50 to 200 mg/m.sup.2, at least once in the cycle.

20. A pharmaceutical composition comprising the immunoconjugate of any of claims 1 to 14, and folinic acid, 5-fluoro-uracil and oxaliplatin.

21. A kit comprising (i) a pharmaceutical composition of the immunoconjugate of any of claims 1 to 14 and (ii) one or more pharmaceutical compositions comprising folinic acid, 5-fluoro-uracil and oxaliplatin, in separate or combined formulations.

22. The pharmaceutical composition according to claim 20 or the kit according to claim 21 for the use for treating cancer.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0166] FIG. 1: Activity of immunoconjugate huMAb2-3-SPDB-DM4 and FOLFOX regimen as single agents or in combination against subcutaneous colon patient-derived xenograft (PDX) CR-IGR-0007P PDX in SCID mice. Tumor volume evolution by treatment group. The curves represent medians + or − MAD (Median Absolute Deviation) at each day for each group.

[0167] FIG. 2: Activity of immunoconjugate huMAb2-3-SPDB-DM4 and FOLFOX regimen as single agents or in combination against subcutaneous colon patient-derived xenograft CR-IGR-0011C PDX, in SCID mice. Tumor volume evolution by treatment group. The curves represent medians + or − MAD at each day for each group.

EXAMPLES

Example 1: Activity of Immunoconjugate huMAb2-3-SPDB-DM4 in Combination with FOLFOX Against Two Subcutaneous Colon Patient-Derived Xenografts CR-IGR-0007P PDX and CR-IGR-0011C PDX in SCID Mice

[0168] Experimental Procedure

[0169] The activity of huMAb2-3-SPDB-DM4 and FOLFOX regimen was evaluated as single agent or in combination in two subcutaneous colon patient-derived xenografts (PDX) (CR-IGR-0007P PDX and CR-IGR-0011C PDX) implanted s.c. in female SCID mice. Control groups were left untreated. The doses of the compounds used are given in mg/kg.

[0170] For the CR-IGR-0007P PDX, treatments were initiated on day 26 post tumour implantation when median tumour burden reached 166.0 mm.sup.3. huMAb2-3-SPDB-DM4 was administered at 5 mg/kg following 3 weekly cycles of IV administrations on days 26, 33 and 40. The FOLFOX regimen was administered following 3 weekly cycles and consisted of IV administrations of folinic acid at 30 mg/kg and oxaliplatin at 5 mg/kg on days 26, 33, and 40 and IV administrations of 5-FU at 28 mg/kg on days 27, 34, and 41.

[0171] For the CR-IGR-0011C PDX, treatments were initiated on day 19 post tumour implantation when median tumour burden reached 123.5 mm.sup.3. huMAb2-3-SPDB-DM4 was administered at 5 mg/kg following 3 weekly cycles of IV administrations on days 19, 26 and 33. FOLFOX regimen were administered following 3 weekly cycles and consisted of IV administrations of folinic acid at 30 mg/kg and oxaliplatin at 5 mg/kg on days 19, 26, and 33 and IV administrations of 5-FU at 28 mg/kg on days 20, 27, and 34.

[0172] For the evaluation of anti-tumor activity, animals were weighed daily and tumors were measured 2 times weekly by caliper. A dosage producing a 20% weight loss at nadir (mean of group) or 10% or more drug deaths, was considered an excessively toxic dosage. Animal body weights included the tumor weights. Tumor volume were calculated using the formula mass (mm.sup.3)=[length (mm)×width (mm)×width (mm)]/2. The primary efficacy end points are ΔT/ΔC, percent median regression, partial and complete regressions (PR and CR).

[0173] Changes in tumor volume for each treated (T) and control (C) are calculated for each tumor by subtracting the tumor volume on the day of first treatment (staging day) from the tumor volume on the specified observation day. The median ΔT is calculated for the treated group and the median ΔC is calculated for the control group. Then the ratio ΔT/ΔC is calculated and expressed as a percentage: ΔT/ΔC=(delta T/delta C)×100.

[0174] The dose is considered as therapeutically active when ΔT/ΔC is lower than 40% and very active when ΔT/ΔC is lower than 10%. If ΔT/ΔC is lower than 0, the dose is considered as highly active and the percentage of regression is dated (Plowman J, Dykes D J, Hollingshead M, Simpson-Herren L and Alley M C. Human tumor xenograft models in NCI drug development. In: Feibig H H B A, editor. Basel: Karger.; 1999 p 101-125):

[0175] % tumor regression is defined as the % of tumor volume decrease in the treated group at a specified observation day compared to its volume on the first day of first treatment.

[0176] At a specific time point and for each animal, % regression is calculated. The median % regression is then calculated for the group:

[00001]$\%\mathrm{regression}\left(\mathrm{at}t\right)=\frac{{\mathrm{volume}}_{t0}-{\mathrm{volume}}_t}{{\mathrm{volume}}_{t0}}\times 100$

[0177] Partial regression (PR): Regressions are defined as partial if the tumor volume decreases to 50% of the tumor volume at the start of treatment.

[0178] Complete regression (CR): Complete regression is achieved when tumor volume=0 mm.sup.3 (CR is considered when tumor volume cannot be recorded).

[0179] Results

[0180] The results for the CR-IGR-0007P PDX are presented on FIG. 1 and Table 1 (below).

[0181] One mouse of control group was found dead on D54; the CR-IGR-0007P is an aggressive tumor and can be cachexic. huMAb2-3-SPDB-DM4 was administered at doses lower than maximal tolerated dose (MTD) and treatments were well tolerated and did not induce toxicity. The FOLFOX regimen was administered at its respective MTD determined in mice non-bearing tumor. In these mice bearing CR-IGR-0007P tumor, cytotoxic treatments were tolerated alone or in combination with body weight loss between 8.1 to 10.8%.

[0182] The huMAb2-3-SPDB-DM4 as single was inactive with a ΔT/ΔC on D49 equal to 76%. The FOLFOX regimen as single agent was active with a ΔT/ΔC equal to 15% (p <0.0001).

[0183] The combination of the huMAb2-3-SPDB-DM4 and FOLFOX regimen was highly active with a ΔT/ΔC inferior to 0% (p<0.0001), a tumor regression of 9% and 1 PR (partial regression). The effect of the combination of huMAb2-3-SPDB-DM4 with FOLFOX was significantly different from the effect of huMAb2-3-SPDB-DM4 alone from day 36 to day 57 and significantly different from the effect of FOLFOX alone from day 44 to 57.

[0184] In conclusion in the CR-IGR-0007P PDX, huMAb2-3-SPDB-DM4 after 3 weekly IV administrations at 5 mg/kg was inactive as single agent, however the FOLFOX regimen was active and the treatment was well tolerated. The combination of the huMAb2-3-SPDB-DM4 and FOLFOX regimen was more active than the single agents.

TABLE-US-00001 TABLE 1 Activity of huMAb2-3-SPDB-DM4 and FOLFOX regimen alone or in combination against subcutaneous colon Patient-Derived-Xenograft, CR-IGR-0007P in SCID mice Dosage in Route mg/kg Drug Mean body Median Median (Dosage (total death weight change ΔT/ΔC % of Biosatitic in cumulated Schedule (day of in % at nadir in % regression Regression p value.sup.a Biological Agent mL/kg) dose) in day death) (day of nadir) (D49) (D49) PR CR (D49) comments Oxaliplatin IV (5)   5 (15) 26, 33, 40 0/6 −9.4 (45)   15 — 0/6 0/6 <000.1 Active Leucovorine IV (5)  30 (90) 26, 33, 40 5-FU IV (5)  28 (84) 27, 34, 41 huMAb2-3- IV (10)  5 (15) 26, 33, 40 0/6 −3.4 (54)   76 — 0/6 0/6 0.1068 Inactive SPDB-DM4 Oxaliplatin IV (5)   5 (15) 26, 33, 40 0/6 −7.6 (43)  <0 9 1/6 0/6 <0.0001 Highly Leucovorine IV (5)  30 (90) 26, 33, 40 active 5-FU IV (5)  28 (84) 27, 34, 41 huMAb2-3- IV (10)  5 (15) 26, 33, 40 SPDB-DM4 Control — — — 0/6 −7.0 (57) — — — — — — .sup.aStatistical analysis. The p-values were obtained using a contrast analysis to compare each treated group versus control using Bonferroni-Holm adjustment for multiplicity after a two-way Anova-Type with repeated measures on tumor volume changes from baseline. A probability less than 5% (p < 0.05) was considered as significant. ΔT/ΔC = ratio of medians of tumor volume changes from baseline between treated and control groups; PR = Partial regression; CR = Complete regression

[0185] The results for the CR-IGR-0011C PDX are presented on FIG. 2 and Table 2 (below).

[0186] Mice of control group exhibited negative body weight changes (nadir of −6.7% on Day 32); the CR-IGR-0011C is an aggressive tumor and can be cachexic. huMAb2-3-SPDB-DM4 was administered at doses lower than maximal tolerated dose (MTD) and treatments were well tolerated and did not induce toxicity.

[0187] The FOLFOX regimen was administered at its MTD determined in mice non-bearing tumor. In these mice bearing CR-IGR-0011C tumor that induced body weight loss, cytotoxic treatments induced additive body weight loss alone or in combination and high calorie dietary supplement for laboratory rodents was added for each group on D24.

[0188] The FOLFOX regimen alone or in combination induced body weight loss between 5.6 to 9.8%, excepted for the group treated by the combination of FOLFOX and huMAb2-3-SPDB-DM4 in which a single mouse lost progressively body weight until reaching more than 20% of body weight loss and died on D24. This death is due to the addition of the tumor aggressiveness and the treatment impact and, considering the entire group, it was not observed a cumulative toxicity with the combination.

[0189] The huMAb2-3-SPDB-DM4 as single agent was highly active with a ΔT/ΔC on D35 inferior to 0% (p<0.0001), a tumor regression of 29% and 2 PR.

[0190] The FOLFOX regimen as single agent was inactive with a ΔT/ΔC equal to 71% (NS).

[0191] The combination of huMAb2-3-SPDB-DM4 and FOLFOX regimen was highly active with a ΔT/ΔC inferior to 0% (p<0.0001), a tumor regression of 76%, 5 PR and 1 CR (complete regression). The effect of the combination of huMAb2-3-SPDB-DM4 with FOLFOX was significantly different from the effect of huMAb2-3-SPDB-DM4 alone from day 22 to day 35 and significantly different from the effect of FOLFOX alone from day 22 to 35.

[0192] In conclusion, in the CR-IGR-0001C PDX, huMAb2-3-SPDB-DM4 after 3 weekly IV administrations at 5 mg/kg was highly active as single agent. FOLFOX was inactive as single agent. The combination of HUMAB2-3-SPDB-DM4 with FOLFOX were significantly more active than the corresponding single agents.

TABLE-US-00002 TABLE 2 Activity of HUMAB2-3-SPDB-DM4 and FOLFOX regimen alone or in combination against subcutaneous colon Patient-Derived-Xenograft, CR-IGR-0011C in SCID mice Dosage in Route mg/kg Drug Mean body Median Median (Dosage (total death weight change ΔT/ΔC % of Biostatitic in cumulated Schedule (day of in % at nadir in % regression Regression p value.sup.a Biological Agent mL/kg) dose) in day death) (day of nadir) (D35) (D35) PR CR (D35) comments Oxaliplatin IV (5)   5 (15) 19, 26, 33 0/6 −9.1 (33)   71 — 0/6 0/6   0.3041 Inactive Leucovorine IV (5)  30 (90) 19, 26, 33 5-FU IV (5)  28 (84) 20, 27, 34 HUMAB2-3- IV (10)  5 (15) 19, 26, 33 0/6 −6.2 (25)  <0 29 2/6 0/6 <0.0001 Highly SPDB-DM4 active Oxaliplatin IV (5)   5 (15) 19, 26, 33 1/6 −6.8 (23)  <0 76 5/6 1/6 <0.0001 Highly Leucovorine IV (5)  30 (90) 19, 26, 33 (D24) active 5-FU IV (5)  28 (84) 20, 27, 34 HUMAB2-3- IV (10)  5 (15) 19, 26, 33 SPDB-DM4 Control — — — 0/6 −6.7 (32) — — — — — — .sup.aStatistical analysis. The p-values were obtained using a contrast analysis to compare each treated group versus control using Bonferroni-Holm adjustment for multiplicity after a two-way Anova-Type with repeated measures on tumor volume changes from baseline. A probability less than 5% (p < 0.05) was considered as significant. ΔT/ΔC = ratio of medians of tumor volume changes from baseline between treated and control groups; PR = Partial regression; CR = Complete regression