ANTITUMOR COMBINATIONS CONTAINING ANTI-CEACAM5 ANTIBODY CONJUGATES, TRIFLURIDINE AND TIPIRACIL

Abstract

The present invention concerns antibody-conjugates comprising an anti-CEACAM5-antibody for use for treating cancer in combination with trifluoridine and tipiracil (TAS-102). The invention further relates to pharmaceutical compositions and kit-of-parts comprising an anti-CEACAM5-antibody in combination with trifluoridine and tipiracil (TAS-102) for use for treating cancer.

Claims

1. An immunoconjugate comprising an anti-CEACAM5-antibody for use for treating cancer in combination with trifluoridine and tipiracil (TAS-102).

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, and stomach cancer.

15. The immunoconjugate for the use of any of claims 1 to 14, wherein the immunoconjugate and TAS-102 are administered simultaneously to a subject in need thereof.

16. The immunoconjugate for the use of claim 15, wherein the immunoconjugate and TAS-102 are formulated (i) in a single pharmaceutical composition comprising the immunoconjugate and TAS-102, 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 trifluoridine and tipiracil, in separate or combined formulations.

17. The immunoconjugate for the use of any of claims 1 to 14, wherein the immunoconjugate and TAS-102 are administered separately or sequentially to a subject in need thereof.

18. The immunoconjugate for the use of claim 17, wherein the immunoconjugate and TAS-102 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 trifluoridine and tipiracil, 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 trifluoridine and tipiracil (TAS-102) are administered in 3 to 6 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 TAS-102 at a dose of from 10 to 100 mg/m.sup.2, wherein TAS-102 comprises trifluoridine and tipiracil in a molar ratio of from 1:0.4 to 1:0.6, at least once in the cycle.

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

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 trifluoridine and tipiracil, 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

[0164] FIG. 1: Activity of immunoconjugate huMAb2-3-SPDB-DM4 and TAS-102 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 TAS-102 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 TAS-102 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 TAS-102 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. A TAS-102 solution was prepared comprising trifluoridine and tipiracil in a molar ratio of 1:0.5. The TAS-102 regimen was administered by oral route at 100 mg/kg/day (expressed as trifluoridine dose) twice a day at approx. 6-hour intervals on days 26 to 30 and on days 33 to 37.

[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. A TAS-102 solution was prepared comprising trifluoridine and tipiracil in a molar ratio of 1:0.5. The TAS-102 regimen was administered by oral route at 100 mg/kg/day (expressed as trifluoridine dose) twice a day at approx. 6-hour intervals on days 19 to 23 and on days 26 to 30.

[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 M C. Human tumor xenograft models in NCI drug development. In: Feibig H H B A, 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]$\%\mathrm{regresstion}\left(\mathrm{at}t\right)=\frac{{\mathrm{volume}}_{t0}-{\mathrm{volume}}_t}{{\mathrm{volume}}_{t0}}\times 100$

[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 PDX 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 TAS-102 regimen was administered at its MTD determined in mice non-bearing tumor. In these mice bearing CR-IGR-0007P PDX tumor, cytotoxic treatments were tolerated alone or in combination with body weight loss between 8.1 to 10.8%.

[0180] The huMAb2-3-SPDB-DM4 as a single agent was inactive with a ΔT/ΔC on D49 equal to 76%. The TAS-102 regimen as single agent was inactive with a ΔT/ΔC equal to 42%.

[0181] The combined huMAb2-3-SPDB-DM4 and TAS-102 regimen was very active with a ΔT/ΔC equal to 9% (p<0.0001). The effect of the combination of huMAb2-3-SPDB-DM4 with TAS-102 was significantly different from the effect of huMAb2-3-SPDB-DM4 alone from day 40 to day 62 and significantly different from the effect of TAS-102 alone from day 49 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. The TAS-102 regimen alone was also inactive and the treatment was tolerated. The combination of the huMAb2-3-SPDB-DM4 and TAS-102 regimen was significantly more active than the single agents.

TABLE-US-00001 TABLE 1 Activity of huMAb2-3-SPDB-DM4 and TAS-102 regimen in combination against subcutaneous colon Patient-Derived-Xenograft, CR-IGR-0007P in SCID mice Dosage in mg/kg Drug Mean body Route (total death weight change Median Median % of Biosatitic (Dosage cumulated Schedule in (day of in % at nadir ΔT/ΔC in regression Regression p value.sup.a Biological Agent in mL/kg) dose) day death) (day of nadir) % (D49) (D49) PR CR (D49) comments TAS-102 PO (10) 100 (1000) BID .sup. 0/6.sup.b −8.1 (31) 42 — 0/6 0/6 0.0356 Inactive 26-30, 33-37 huMAb2-3-  IV (10) 5 (15) 26, 33, 40 0/6 −3.4 (54) 76 — 0/6 0/6 0.1068 Inactive SPDB-DM4 TAS-102 PO (10) 100 (1000) BID 0/6 −10.8 (39)  6 — 0/6 0/6 <0.0001 Very active huMAb2-3-  IV (10) 5 (15) 26-30, 33-37 SPDB-DM4 26, 33, 40 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 PDX 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.

[0185] The TAS-102 regimen was administered at its MTD determined in mice non-bearing tumor. In these mice bearing CR-IGR-0011C PDX 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 TAS-102 regimen alone or in combination induced body weight loss superior to 20% and death on D34 in the group treated with TAS-102 alone.

[0186] 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).

[0187] The TAS-102 regimen as single agent was very active with a ΔT/ΔC equal to 29% (p<0.0027).

[0188] The combination of huMAb2-3-SPDB-DM4 and TAS-102 regimen was highly active with a ΔT/ΔC inferior to 0% (p<0.0001), a tumor regression of 83%, 4 PR and 2 CR (complete regression). The effect of the combination of huMAb2-3-SPDB-DM4 1 with TAS-102 was significantly different from the effect of huMAb2-3-SPDB-DM4 alone from day 27 to day 33 and significantly different from the effect of TAS-102 alone from day 30 to 35.

[0189] 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. TAS-102 was also active as single agent. The combination of HUMAB2-3-SPDB-DM4 with TAS-102 was significantly more active than single agents.

TABLE-US-00002 TABLE 2 Activity of HUMAB2-3-SPDB-DM4 and TAS-102 regimen in combination against subcutaneous colon Patient-Derived-Xenograft, CR-IGR-0011C in SCID mice Dosage in mg/kg Drug Mean body Biosta- Route (total death weight change Median Median % of tistic (Dosage cumulated Schedule in (day of in % at nadir ΔT/ΔC in regression Regression p value.sup.a Biological Agent in mL/kg) dose) day death) (day of nadir) % (D35) (D35) PR CR (D35) comments TAS-102 PO (10) 100 (1000) BID 1/6 (D34) −21.1 (33) 29 — 0/6 0/6 0.0027 Active 19-23, 26-30 Toxic 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 TAS-102 PO (10) 100 (1000) BID 0/6 −20.2 (33) <0 83 4/6 2/6 <0.0001 Highly HUMAB2-3-  IV (10) 5 (15) 19-23, 26-30 active SPDB-DM4 19, 26, 33 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