ANTITUMOR COMBINATIONS CONTAINING ANTI-CEACAM5 ANTIBODY CONJUGATES AND CETUXIMAB

Abstract

The present invention concerns antibody-conjugates comprising an anti-CEACAM5-antibody for use for treating cancer in combination with cetuximab. The invention further relates to pharmaceutical compositions and kit-of-parts comprising an anti-CEA-CAM5-antibody in combination with cetuximab for use for treating cancer.

Claims

1. An immunoconjugate comprising an anti-CEACAM5-antibody for use for treating cancer in combination with cetuximab.

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 colorectal cancer.

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

16. The immunoconjugate for the use of claim 15, wherein the immunoconjugate and cetuximab are formulated (i) in a single pharmaceutical composition comprising the immunoconjugate and cetuximab, or (ii) in the form of two separate pharmaceutical compositions, wherein one pharmaceutical composition comprises the immunoconjugate, and on the other pharmaceutical composition comprises cetuximab.

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

18. The immunoconjugate for the use of claim 17, wherein the immunoconjugate and cetuximab are formulated in the form of two separate pharmaceutical compositions, wherein (i) one pharmaceutical composition comprises the immunoconjugate, and (ii) the other pharmaceutical composition comprises cetuximab.

19. The immunoconjugate for the use of any of claims 1 to 18, wherein the immunoconjugate comprising an anti-CEACAM5-antibody, and cetuximab 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 a cycle; administering cetuximab at a dosage of from 100 to 600 mg/m.sup.2 at least once in a cycle.

20. A pharmaceutical composition comprising the immunoconjugate of any of claims 1 to 14, and cetuximab.

21. A kit comprising (i) a pharmaceutical composition of the immunoconjugate of any of claims 1 to 14 and (ii) a pharmaceutical composition comprising cetuximab.

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

[0164] FIG. 1: Activity of immunoconjugate huMAb2-3-SPDB-DM4 and cetuximab 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.

[0165] FIG. 2: Activity of immunoconjugate huMAb2-3-SPDB-DM4 and cetuximab 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 Cetuximab Against Two Subcutaneous Colon Patient-Derived Xenografts CR-IGR-0007P PDX and CR-IGR-0011C PDX in SCID Mice

[0166] Experimental Procedure

[0167] The activity of huMAb2-3-SPDB-DM4 and cetuximab 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.

[0168] 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. Cetuximab was administered at 10 mg/day as IV administrations on days 26, 33, 40, and 44.

[0169] 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. Cetuximab was administered at 10 mg/kg as IV administrations on days 19, 23, 26, 30 and 33.

[0170] 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).

[0171] 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.

[0172] 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 MC. Human tumor xenograft models in NCI drug development. In: Feibig HH BA, editor. Basel: Karger.; 1999 p 101-125):

[0173] % 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.

[0174] At a specific time point and for each animal, % regression is calculated. The median % regression is then calculated for the group:

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

[0175] Partial regression (PR): Regressions are defined as partial if the tumor volume decreases to 50% of the tumor volume at the start of treatment.

[0176] Complete regression (CR): Complete regression is achieved when tumor volume=0 mm.sup.3 (CR is considered when tumor volume cannot be recorded).

[0177] Results

[0178] The results for the CR-IGR-0007P PDX are presented on FIG. 1 and Table 1 (below).

[0179] 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 and cetuximab were administered at doses lower than maximal tolerated dose (MTD) and treatments were well tolerated and did not induce toxicity.

[0180] The huMAb2-3-SPDB-DM4 as a single agent was inactive with a ΔT/ΔC on D49 equal to 76%. The cetuximab regimen as single agent was very active with a ΔT/ΔC equal to 2% (p<0.0001).

[0181] The combined huMAb2-3-SPDB-DM4 and cetuximab regimen was ΔT/ΔC inferior to 0% (p<0.0001), a tumor regression of 67% and 6 PR (partial regression). The effect of the combination of huMAb2-3-SPDB-DM4 with cetuximab was significantly different from the effect of cetuximab alone from day 36 to day 62 and significantly different from the effect of huMAb2-3-SPDB-DM4 alone from day 33 to 62.

[0182] 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. Cetuximab regimen was very active and the treatment was well tolerated. The combination of the huMAb2-3-SPDB-DM4 and cetuximab regimen was more active than the single agents.

TABLE-US-00001 TABLE 1 Activity of huMAb2-3-SPDB-DM4 and cetuximab in combination against subcutaneous colon Patient-Derived-Xenograft, CR-IGR-0007P in SCID mice Dosage in Route mg/kg Drug Mean body Median (Dosage (total death weight change Median % of Biosatitic in mL/ cumulated Schedule in (day of in % at nadir ΔT/ΔC in regression Regression p value.sup.a Biological Agent kg) dose) day death) (day of nadir) % (D49) (D49) PR CR (D49) comments Cetuximab IV (10) 10 (40) 26, 33, 40, 44 0/6 −4.2 (45) 2 — 0/6 0/6 <0.0001 Very active huMAb2-3- IV (10)  5 (15) 26, 33, 40 0/6 −3.4 (54) 76 — 0/6 0/6   0.1068 Inactive SPDB-DM4 Cetuximab IV (10) 10 (40) 26, 33 ,40, 44 0/6 −4.7 (40) <0 67 6/6 0/6 <0.0001 Very active 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

[0183] The results for the CR-IGR-0011C PDX are presented on FIG. 2 and Table 2 (below).

[0184] 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 and cetuximab were administered at doses lower than maximal tolerated dose (MTD) and treatments were well tolerated and did not induce toxicity.

[0185] 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 (partial regression). The cetuximab regimen as single agent inactive with a ΔT/ΔC equal to 44% (NS).

[0186] The combination of huMAb2-3-SPDB-DM4 and cetuximab regimen was highly active with a ΔT/ΔC inferior to 0% (p<0.0001), a tumor regression of 52% and 3 PR. The effect of the combination of huMAb2-3-SPDB-DM4 with cetuximab was significantly different from the effect of huMAb2-3-SPDB-DM4 alone from day 30 to day 33 and significantly different from the effect of cetuximab alone from day 30 to 35.

[0187] 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. Cetuximab was inactive as single agent. The combination of HUMAB2-3-SPDB-DM4 with cetuximab was significantly more active than the single agents.

TABLE-US-00002 TABLE 2 Activity of HUMAB2-3-SPDB-DM4 and cetuximab in combination against subcutaneous colon Patient-Derived-Xenograft, CR-IGR-0011C in SCID mice Dosage in Route mg/kg Drug Mean body Median (Dosage (total death weight change Median % of Biosatitic in mL/ cumulated Schedule in (day of in % at nadir ΔT/ΔC in regression Regression p value.sup.a Biological Agent kg) dose) day death) (day of nadir) % (D35) (D35) PR CR (D35) comments Cetuximab IV (10) 10 (50) 19, 23, 26, 30, 33 0/6 −3.9 (33) 44 — 0/6 0/6   0.1528 Inactive 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 Cetuximab IV (10) 10 (40) 19, 23, 26, 30, 33 0/6 −2.9 (24) <0 52 3/6 0/6 <0.0001 Highly HUMAB2-3- IV (10)  5 (15) 19, 26, 33 active 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