ANTITUMOR COMBINATIONS CONTAINING ANTI-CEACAM5 ANTIBODY CONJUGATES AND FOLFIRI

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 irinotecan (FOLFIRI). 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 irinotecan (FOLFIRI) 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 irinotecan (FOLFIRI).

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 FOLFIRI are administered simultaneously to a subject in need thereof.

16. The immunoconjugate for the use of claim 15, wherein the immunoconjugate and FOLFIRI are formulated (i) in a single pharmaceutical composition comprising the immunoconjugate and FOLFIRI, 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 irinotecan, in separate or combined formulations.

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

18. The immunoconjugate for the use of claim 17, wherein the immunoconjugate and FOLFIRI 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 irinotecan, 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 irinotecan (FOLFIRI) are administered in 8 to 16 cycles, wherein one cycle comprises: administering the immunoconjugate at a dosage of from 60 to 210 mg/m.sup.2, at least once in the cycle; administering folinic acid at a dosage 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 dosage of from 1000 to 2000 mg/m.sup.2, at least once in the cycle, and administering irinotecan at a dosage of from 100 to 300 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 irinotecan.

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 irinotecan, 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

[0168] FIG. 1: Activity of immunoconjugate huMAb2-3-SPDB-DM4 and FOLFIRI 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.

[0169] FIG. 2: Activity of immunoconjugate huMAb2-3-SPDB-DM4 and FOLFIRI 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 FOLFIRI Against Two Subcutaneous Colon Patient-Derived Xenografts CR-IGR-0007P PDX and CR-IGR-0011C PDX in SCID Mice

[0170] Experimental Procedure

[0171] The activity of huMAb2-3-SPDB-DM4 and FOLFIRI 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.

[0172] 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 FOLFIRI regimen was administered following 3 weekly cycles and consisted of IV administrations of folinic acid at 60 mg/kg and irinotecan at 22 mg/kg on days 26, 33, and 40 and IV administrations of 5-FU at 56 mg/kg on days 27, 34, and 41.

[0173] 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. FOLFIRI regimen were administered following 3 weekly cycles and consisted of IV administrations of folinic acid at 60 mg/kg and irinotecan at 22 mg/kg on days 19, 26, and 33 and IV administrations of 5-FU at 56 mg/kg on days 20, 27, and 34.

[0174] 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).

[0175] 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.

[0176] 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 BA, editor. Basel: Karger.; 1999 p 101-125):

[0177] % 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.

[0178] At a specific time point and for each animal, % regression is calculated. The median % regression is then calculated for the group:

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

[0179] Partial regression (PR): Regressions are defined as partial if the tumor volume decreases to 50% of the tumor volume at the start of treatment.

[0180] Complete regression (CR): Complete regression is achieved when tumor volume=0 mm.sup.3 (CR is considered when tumor volume cannot be recorded).

[0181] Results

[0182] The results for the CR-IGR-0007P PDX are presented on FIG. 1 and Table 1 (below).

[0183] 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 FOLFIRI 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%, with the exception of one mouse in the group treated with the combination, which lost progressively body weight until reaching more than 20% of body weight loss and death on D48.

[0184] The huMAb2-3-SPDB-DM4 as a single agent was inactive with a ΔT/©C on D49 equal to 76%. The FOLFIRI regimen as single agent was highly active with a ΔT/ΔC inferior to 0% (p<0.0001) and a tumor regression of 18% (Table 1).

[0185] The combined huMAb2-3-SPDB-DM4 and FOLFIRI regimen was highly active with a ΔT/ΔC inferior to 0% (p<0.0001), a tumor regression of 47% and 4 PR (partial regression). The effect of the combination of huMAb2-3-SPDB-DM4 with FOLFIRI was significantly different from the effect of huMAb2-3-SPDB-DM4 alone from day 33 to day 62 and significantly different from the effect of FOLFIRI alone from day 33 to 62.

[0186] 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 FOLFIRI regimen was highly active and the treatment was well tolerated. The combination of the huMAb2 SPDB-DM4 and FOLFIRI regimen was more active than the single agents.

TABLE-US-00001 TABLE 1 Activity of huMAb2-3-SPDB-DM4 and FOLFIRI regimen in combination against subcutaneous colon Patient-Derived- Xenograft, CR-IGR-0007P in SCID mice Mean body Dosage in weight Median Route mg/kg Drug change Median % (Dosage (total Schedule death in % at ΔT/ΔC of Biosatitic in cumulated in (day of nadir (day in % regression Regression p value.sup.a Biological Agent mL/kg) dose) day death) of nadir) (D49) (D49) PR CR (D49) comments Irinotecan IV (10) 22 (66) 26, 33, 40 0/6.sup.b −8.9 (46) <0 18 0/6 0/6 <0.0001 Highly Folinic acid IV (5) 60 (180) 26, 33, 40 active 5-FU IV (5) 56 (168) 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 Irinotecan IV (10) 22 (66) 26, 33, 40 1/6  −9.8 (45) <0 47 4/6 0/6 <0.0001 Highly Folinic acid IV (5) 60 (180) 26, 33, 40 (D48) active 5-FU IV (5) 56 (168) 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

[0187] The results for the CR-IGR-0011C PDX are presented on FIG. 2 and Table 2 (below).

[0188] 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.

[0189] The FOLFIRI 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. The FOLFIRI regimen alone or in combination induced body weight loss between 5.6 to 9.8%.

[0190] 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. The FOLFIRI regimen as single agent was very active with a ΔT/ΔC equal to 2% (p<0.0001).

[0191] The combination of huMAb2-3-SPDB-DM4 and FOLFIRI regimen was highly active with a ΔT/ΔC inferior to 0% (p<0.0001), a tumor regression of 88%, 6 PR and 2 CR (complete regression). The effect of the combination of huMAb2-3-SPDB-DM4 with FOLFIRI 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 FOLFIRI alone from day 30 to 35. 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. FOLFIRI was also very active as single agent. The combination of HUMAB2-3-SPDB-DM4 with FOLFIRI was significantly more active than the corresponding single agents.

TABLE-US-00002 TABLE 2 Activity of HUMAB2-3-SPDB-DM4 and FOLFIRI regimen in combination against subcutaneous colon Patient-Derived- Xenograft, CR-IGR-0011C in SCID mice Mean body Dosage in weight Route mg/kg Drug change Median Median (Dosage (total Schedule death in % at ΔT/ΔC % of Biostatistic in cumulated in (day of nadir (day in % regression Regression p value.sup.a Biological Agent mL/kg) dose) day death) of nadir) (D35) (D35) PR CR (D35) comments Irinotecan IV (10) 22 (66) 19, 26, 33 0/6 −5.6 (24) 2 — 0/6 0/6 <.0.0001 Very Folinic acid IV (5) 60 (180) 19, 26, 33 active 5-FU IV (5) 56 (168) 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 Irinotecan IV (10) 22 (66) 19, 26, 33 0/6 −7.6 (35) <0 88 6/6 2/6 <0.0001  Highly Folinic acid IV (5) 60 (180) 19, 26, 33 active 5-FU IV (5) 56 (168) 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