(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2010/0160233 A1
`(43) Pub. Date:
`Jun. 24, 2010
`BSSERY et al.
`
`US 2010016O233A1
`
`(54) ANTITUMOUR COMBINATIONS
`CONTAINING AVEGF INHIBITINGAGENT
`AND RINOTECAN
`
`(75) Inventors:
`
`Marie-Christine BISSERY,
`Charenton le Pont (FR); Marielle
`CHIRON-BLONDEL, Paris (FR);
`Pascale LEJEUNE, Vitry (FR):
`Patricia VRIGNAUD, Combs la
`Ville (FR)
`
`Correspondence Address:
`ANDREA Q. RYAN
`SANOF-AVENTIS U.S. LLC
`104.1 ROUTE 202-206, MAIL CODE: D303A
`BRIDGEWATER, NJ 08807 (US)
`
`(73) Assignee:
`
`Aventis Pharma S.A., Antony (FR)
`
`(21) Appl. No.:
`
`12/631,767
`
`(22) Filed:
`
`Jan. 4, 2010
`Related U.S. Application Data
`(63) Continuation of application No. PCT/FR2008/
`000943, filed on Jul. 2, 2008.
`Foreign Application Priority Data
`
`(30)
`
`Jul. 5, 2007 (FR) ....................................... O704868
`Publication Classification
`
`(51) Int. Cl.
`(2006.01)
`A638/16
`(2006.01)
`A63L/4375
`(52) U.S. Cl. ........................................... 514/12: 514/283
`(57)
`ABSTRACT
`Disclosed are antitumor combinations of VEGF inhibitors
`with Irinotecan and the use thereof in the treatment of neo
`plastic diseases.
`
`Regeneron Exhibit 2015
`Page 01 of 05
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`

`US 2010/01 60233 A1
`
`Jun. 24, 2010
`
`ANTITUMIOUR COMBINATIONS
`CONTAINING AVEGF INHIBITINGAGENT
`AND RINOTECAN
`
`0001. The present invention relates to the combinations of
`a VEGF inhibitor and of a chemotoxic agent of the topoi
`Somerase inhibitor class, useful in the treatment of neoplastic
`diseases.
`0002 VEGF inhibitors, which are inhibitors of vascular
`endothelial growth factor, are in the majority of cases biologi
`cal products selected from Soluble receptors, antisenses,
`RNAaptamers and antibodies. The topoisomerase inhibitors
`of use in the treatment of known neoplastic diseases are
`selected from camptothecins, including CPT 11, topotecan
`and pyridobenzoindole. The present combination is in par
`ticular directed toward the treatment of colon cancer or of
`stomach cancer.
`0003. The description and the preparation of the VEGF
`inhibitor preferably used in the invention, which is a VEGF
`Trap chimeric protein, is described in patent application WO
`00/75319. There are several embodiments of the chimeric
`protein.
`0004. The embodiment corresponding to VEGF Trap is
`the one described in FIG.24 (sequence) of WO00/75319. The
`VEGF Trap used in the invention is a fusion protein compris
`ing the signal sequence of VEGFR1 fused to the D2 Ig domain
`of the VEGFR1 receptor, itself fused to the D3 Ig domain of
`the VEGFR2 receptor, in turnfused to the Fc domain of IgG1,
`also known as VEGFR1R2-FcAC1 or Flt1D2. Flk1D3.
`FCAC1.
`0005. In general, the doses of VEGF Trap used in humans,
`which depend on factors specific to the individual to be
`treated, are between 1 and 10 milligrams per kilo when the
`administration is carried out Subcutaneously or intravenously.
`0006 Among the topoisomerase inhibitors, irinotecan,
`also known under the nonproprietary name CPT-11, is pref
`erably used.
`0007 Irinotecan is generally used intravenously at a dose
`of between 100 mg/m and 500 mg/m depending on the
`administration scheme. A dose of 150 mg/m is, for example,
`used for a weekly scheme and a dose of between 200 and 400
`mg/m is, for example, used for a scheme every three weeks.
`0008. An article by H Hurwitz, L. Fehrenbacher, W
`Novotny, T Cartwright, J Hainsworth, W Heim, J Berlin, A
`Baron, S Griffing, E Holmgren, NFerrara, G Fyfe, B Rogers,
`R Ross, F Kabbinavar, published in “The New England Jour
`nal of Medicine” has described a clinical trial proving a better
`survival rate when the combination of bevacizumab with
`irinotecan, 5FU and leucovorin is used, compared with the
`same combination not containing bevacizumab. In this clini
`cal trial, there is nothing to prove that the improvement in the
`survival rate comes from the combination of irinotecan with
`bevacizumab, it may just as easily come from the combina
`tion of 5FU or of leucovorin with bevacizumab, or may come
`from the quadruple combination. Now, since it is known that
`each of the anticancer agents brings toxic side effects, along
`with its therapeutic effect, it appears to be advisable to limit
`their presence as much as possible, especially when the same
`effect can be obtained in the absence of at least one of them.
`Furthermore, this article does not provide evidence of any
`synergistic effect within the meaning of Corbett, i.e. an effect
`which cannot be obtained with each of the elements of the
`combination used alone at its maximum tolerated dose.
`
`0009 VEGF Trap is a soluble receptor created by fusion of
`the second Ig domain of VEGFR-1 with the third Ig domain of
`VEGFR-2, which is subsequently fused to the Fc part of a
`human IgG1. Like the VEGFR-1 receptor, aflibercept (VEGF
`Trap) has a very high affinity for VEGF-A, with a Kd of 0.5
`picoM. The high-affinity binding of VEGF Trap with
`VEGF-A results in the formation of a complex which pre
`vents VEGF from binding to and activating its receptors at the
`surface of cells.
`0010. In comparison with Avastin (or bevacizumab),
`VEGF Trap is a soluble receptor, whereas Avastin is an anti
`body directed against VEGF-A. VEGF Trap has a much
`higher affinity for VEGF-A than that of Avastin, and a differ
`ent selectivity profile since VEGF Trap also binds to the other
`ligands of VEGFR1-2 receptors, i.e. to PIGF (placental
`growth factor) and to VEGF-B. Furthermore, VEGF Trap has
`a molecular weight which is substantially less than that of
`Avastin (115 kDa for aflibercept versus 160 kDa for Avastin),
`more favorable to penetration in solid tumors.
`0011. It has now been found, and it is this which is the
`subject of the present invention, that the effectiveness of
`VEGF inhibitors can be considerably improved when they are
`administered in combination with at least one Substance of
`therapeutic use in anticancer treatments which has a mecha
`nism of action different than that of VEGF inhibitors.
`0012 Moreover, since the activity of the products depends
`on the doses used, it is possible to use higher doses and to
`increase the activity by reducing the toxicity phenomena or
`by delaying their appearance, through the combining with the
`VEGF inhibitors or with their analogs of other therapeutically
`active Substances of growth factors of hematopoietic type,
`such as G-CSF or GM-CSF, or certain interleukins.
`0013 More particularly, the invention relates to the com
`binations of VEGF Trap with irinotecan.
`0014. The improved effectiveness of a combination
`according to the invention can be demonstrated by determin
`ing the therapeutic synergism.
`0015. A combination shows atherapeutic synergism if it is
`therapeutically superior to both the constituents used at the
`optimum dose thereof.
`0016. In order to demonstrate the effectiveness of a com
`bination, it may be necessary to compare the maximum tol
`erated dose of the combination with the maximum tolerated
`dose of each of the isolated constituents in the study under
`consideration. This effectiveness can be quantified, for
`example, by the logo cell kill, which is determined according
`to the following equation:
`logo cell kill-T-C(days), 3.32XT
`in which T-C represents the delay in growth of the cells, which
`is the average time, in days, for the tumors of the treated group
`(T) and the tumors of the control group (C) to have reached a
`predetermined value (1 g for example), and T represents the
`time, in days, necessary for the volume of the tumor to double
`in the control animals T. H. Corbett et al., Cancer, 40, 2660.
`2680 (1977); F. M. Schabelet al., Cancer Drug Development,
`Part B. Methods in Cancer Research, 17, 3-51, New York,
`Academic Press Inc. (1979). A product is considered to be
`active if logo cell kill is greater than or equal to 0.7. A product
`is considered to be very active if logo cell kill is greater than
`2.8.
`0017. The combination, used at its own maximum toler
`ated dose, in which each of the constituents is presentata dose
`generally less than or equal to its maximum tolerated dose,
`
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`Page 02 of 05
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`

`

`US 2010/01 60233 A1
`
`Jun. 24, 2010
`
`will show therapeutic synergy when the logo cell kill is
`greater than the value of the logo cell kill of the best con
`stituent when it is administered alone, and in particular has a
`Superiority of at least one log cell kill.
`0.018. The effectiveness of the combinations on solid
`tumors can be determined experimentally in the following
`way:
`0019. The animals subjected to the experiment, generally
`mice, are grafted bilaterally, subcutaneously, with 30 to 60
`mg of an HCT116 human tumor fragment (Brattain, M. G.,
`Fine, W. D., Khaled, F. M., Thompson, J. and Brattain, D. E.,
`Heterogeneity of malignant cells from a human colonic car
`cinoma. Cancer Res., 1981, 41, 1751-1756) on day 0. The
`animals bearing the tumors are randomized before being Sub
`jected to the various treatments and controls. In the case of
`treatment of tumors of the present invention, the tumors were
`allowed to develop to a size of between 48 and 294 mg, which
`made it possible to have a median tumor per group of between
`129 and 162 mg. The animals which underwent the treatment
`with VEGF Trap alone had a weight of between 17.1 and 22.7
`g, the animals having undergone the treatment withirinotecan
`alone had a weight of between 17.5 and 22.3 g, and those
`which received the combination had a weight of between 17.5
`and 23.6 g. Animals bearing tumors were also subjected to the
`same treatments with the excipient alone in order to be able to
`dissociate the toxic effect of the excipient from the actual
`effect of the chemotherapy on the tumor. The chemotherapy
`was begun on day 12 after the tumor graft. The VEGF Trap
`injections were given subcutaneously simultaneously with
`the irinotecan injections, which themselves were given intra
`venously, according to a daily double injection. These injec
`tions were carried out on days 12, 15 and 18 after implantation
`of the tumor. The various groups of animals are weighed three
`to four times per week until the maximum weight loss is
`reached, and then the groups are weighed at least once a week
`until the end of the trial.
`0020. The tumors are measured two or three times a week
`until the tumor reaches approximately 2 g or until the death of
`the animal if the latter occurs before the tumor reaches 2 g.
`The animals are autopsied at the time of sacrifice.
`0021. The antitumor activity is determined according to
`the various parameters recorded.
`0022. By way of examples, the following tables give the
`results obtained with combinations of VEGF Trap and of
`irinotecan used at their optimum dose.
`0023 The present invention also relates to the kits of phar
`maceutical compositions containing the products used in the
`combinations according to the invention.
`0024. The products which constitute the combination may
`be administered simultaneously, separately or sequentially in
`Such a way as to obtain the maximum effectiveness of the
`combination; it being possible for each administration to have
`a variable duration ranging from a rapid total administration
`to a continuous infusion.
`0025. As a result of this, for the purpose of the present
`invention, the combinations are not only limited to those
`which are obtained by physical association of the constitu
`ents, but also to those which allow a separate administration
`that can be simultaneous or sequential.
`0026. The compositions according to the invention are
`preferably compositions that can be administered parenter
`ally.
`0027 Compositions for parenteral administration are gen
`erally pharmaceutically acceptable sterile solutions or Sus
`
`pensions which may optionally be prepared extemporane
`ously at the time of use. For the preparation of nonaqueous
`Solutions or Suspensions, natural plant oils such as olive oil,
`sesame oil or liquid paraffin, or injectable organic esters such
`as ethyl oleate, may be used. Aqueous sterile Solutions may be
`constituted of a solution of the product in water. Aqueous
`Solutions are suitable for intravenous administration insofar
`as the pH is Suitably adjusted and the isotonicity is produced,
`for example, by means of a Sufficient amount of sodium
`chloride or of glucose. The sterilization can be carried out by
`heating or by any other means which does not detrimentally
`alter the composition. The combinations may also be in the
`form of liposomes or in the form of an association with
`Supports such as cyclodextrins or polyethylene glycols.
`0028. In the combinations according to the invention, the
`application of the constituents of which may be simultaneous,
`separate or sequential, it is particularly advantageous for the
`amount of VEGF Trap derivative to represent from 10% to
`80% by weight of the combination, it being possible for this
`content to vary according to the nature of the associated
`substance, to the desired effectiveness and to the nature of the
`cancer to be treated.
`0029. The combinations according to the invention are of
`particular use in the treatment of colon cancer and/or stomach
`cancer. In particular, they can have the advantage of being
`able to use the constituents at doses which are much lower
`than those at which they are used alone.
`0030 The following example illustrates a combination
`according to the invention.
`
`EXAMPLE
`0031. Vials of 1 cm containing 25 mg of VEGF Trap
`which are diluted in a buffer of 5 mM phosphate, 5 mM
`sodium citrate, 100 mM sodium chloride, polysorbate 20 and
`20% sucrose are prepared according to the usual technique,
`for Subcutaneous administration. The administration Volume
`per mouse is 0.1 ml. The VEGF Trap is administered once a
`day, on days 12, 15 and 18 after implantation of the tumor.
`0032 0.3 ml per mouse is prepared, according to the usual
`technique, for intravenous administration, from a commer
`cially available solution at 20 mg/ml of irinotecan to be
`diluted with 5% dextrose in water.
`0033. These solutions are administered simultaneously,
`after a suitable dilution.
`0034. The treatment with irinotecan is repeated twice a
`day, with a 4-hour interval, on days 12, 15 and 18 after
`implantation of the tumor.
`0035. The results of the trial are attached in the appended
`table.
`0036 Tumor doubling time=3.2 days.
`0037 Abbreviations used: (T-C) delay in growth of the
`tumor, Icklog cell kill.
`0038 Toxicity was observed for irinotecan alone at the
`doses of 52.4, 32.5 and 20.2 mg/kg/injection owing to a death
`at the dosage of 52.4 and a weight loss of greater than 20% for
`the two lower dosages. Thus, the maximum tolerated dose for
`irinotecan was 12.5 mg/kg/inj (total injected dose of 75.0
`mg/kg). The dose of 12.5 mg/kg/injection was found to be
`active with an Ick of 1.8.
`0039. For the VEGF Trap, the product was well-tolerated
`at all the dosages tested and was found to be active with an Ick
`of 1.7 at 40 mg/kg/administration and 25 mg/kg/administra
`
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`

`

`US 2010/01 60233 A1
`
`3
`
`Jun. 24, 2010
`
`tion. The lower dose of 10 mg/kg/administration is also
`active, with an Ick of 1.3. The dose of 2.5 mg/kg/administra-
`tion is inactive.
`0040. For the combination ofirinotecan at 32.5 mg/kg/inj,
`whatever the dose of VEGF Trap, the combination was found
`to be toxic with a weight loss of 18% close to toxicity. The
`lower dose of 20.2 mg/kg/inj ofirinotecan with 40 mg/kg of
`VEGF Trap was considered to be the maximum tolerated
`dose. This dose had an Ick of 3.0, judged to be very active. The
`same level of activity was found with the lower doses of
`VEGF Trap, such as 25, 10, 2.5 mg/kg/administration (Ick of
`2.9, 3.0 and 2.9, respectively).
`0041
`Irinotecan at 12.5 mg/kg/inj combined with VEGF
`Trap at 40 mg/kg/administration is active with an Ick of 2.7.
`
`This antitumor activity is maintained with 25 and 10 mg/kg of
`VEGF Trap (Ick of 2.9 and 2.7, respectively). The combina
`tion with 2.5 mg/kg/administration of VEGF Trap has an
`activity of 2.0 Ick.
`0042. In conclusion, the activity of the combination of
`VEGF Trap with irinotecan shows a synergistic effect with a
`log cell kill of 3.0, at the maximum tolerated dose of the
`combination, which corresponds to more than 1 log cell kill
`compared with the activity of each of the compounds used
`alone, which exhibits a log cell kill of 1.8 and 1.7 (for irino
`tecan at 12.5 mg/kg/injection and VEGF Trap at 40 mg/kg/
`administration, respectively). An antitumor activity is main
`tained at several levels of doses below the maximum tolerated
`dose of the combination.
`
`Route of
`adminis-
`tration
`
`i.v.
`0.3 ml
`
`Agent
`Group (alone)
`
`CPT-11
`
`6
`
`7
`
`8
`
`9
`
`2 VEGF
`Trap
`
`S.C.
`0.1 ml
`
`3
`
`4
`
`5
`
`Dosage
`in mg/kg
`Scheme
`per
`injection in days
`
`Death due
`to the
`product,
`with day
`of death
`
`Average
`Weight
`time for the
`loss of
`tumor to
`the mouse
`reach 750
`(date of
`low-point) mg in days
`
`Total
`dose in
`mg/kg
`
`Survivors
`without
`tumor at
`day 144 Comments
`
`Log
`cell
`kill
`
`T-C in
`days
`
`52.4
`
`12, 15, 18
`
`314.4
`
`1/8 (23)
`
`32.5
`
`(2x/d)
`
`195.0
`
`O7
`
`20.2
`
`(4h apart)
`
`121.2
`
`12.5
`
`75.0
`
`40.0
`
`12, 15, 18
`
`120.0
`
`2S.O
`
`1O.O
`
`2.5
`
`75.0
`
`3O.O
`
`7.5
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`— Toxic
`
`— Toxic
`
`— Toxic
`
`Of8
`
`Of8
`
`0.8
`
`HNTD
`active
`HDT
`active
`active
`
`19.1
`
`1.8
`
`18.3
`
`1.7
`
`18.3
`
`1.7
`
`13.S
`
`1.3
`
`1.8
`
`active
`
`37.6
`
`36.8
`
`36.8
`
`32.O
`
`2O2
`
`1.7
`
`O2
`
`O8 inactive
`
`— Toxic
`
`-27.0
`(21)
`-23.1
`(19)
`-21.9
`(19)
`-15.9
`(21)
`-3.1
`(13)
`-2.2
`(13)
`-50
`(19)
`-8.3
`(19)
`-16.2
`(21)
`
`-18.6
`(21)
`-18.2
`(19)
`-18.4
`(19)
`-13.0
`(21)
`-11.9
`(19)
`-13.0
`(19)
`-12.5
`(19)
`-11.0
`(19)
`-11.9
`(19)
`-10.0
`(21)
`-14.3
`(21)
`-9.6
`(25)
`
`— Toxic
`
`— Toxic
`
`— Toxic
`
`SO.9
`
`49.6
`
`50.7
`
`49.7
`
`46.8
`
`49.8
`
`47.0
`
`40.O
`
`18.5
`
`32.4
`
`3.0
`
`31.1
`
`2.9
`
`Of8
`
`HNTD
`highly active
`08 high active
`
`32.2
`
`3.0
`
`08 high active
`
`31.2
`
`2.9
`
`08 high active
`
`28.3
`
`2.7
`
`0.8
`
`active
`
`31.3
`
`2.9
`
`08 high active
`
`28S
`
`2.7
`
`0.8
`
`active
`
`21.5
`
`2.0
`
`0.8
`
`active
`
`Of 10
`
`CPT-11
`
`i.v.
`0.3 ml
`VEGF Trap s.c.
`0.1 ml
`
`O
`
`1
`
`2
`
`3
`
`4
`
`5
`
`6
`
`7
`
`8
`
`9
`
`2O
`
`21
`
`1
`
`Vehicle
`
`S.C.
`0.1 ml
`i.v.
`0.3 ml
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`O.8
`
`95.0
`
`2O.O
`
`95.0
`75.0
`95.0
`3O.O
`95.0
`7.5
`21.2
`2O.O
`21.2
`75.0
`21.2
`3O.O
`21.2
`7.5
`75.0
`2O.O
`75.0
`75.0
`75.0
`3O.O
`75.0
`7.5
`
`32.5
`
`40.0
`
`12, 15, 18
`(2xid)
`12, 15, 18
`
`32.5
`2S.O
`32.5
`1O.O
`32.5
`2.5
`2O2
`40.O
`2O2
`2S.O
`2O2
`1O.O
`2O2
`2.5
`12.5
`40.O
`12.5
`2S.O
`12.5
`1O.O
`12.5
`2.5
`— 12, 15, 18
`
`— 12, 15, 18
`(2xid) (4h
`later)
`
`Regeneron Exhibit 2015
`Page 04 of 05
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`

`

`US 2010/01 60233 A1
`
`Jun. 24, 2010
`
`What is claimed is:
`1. A method of treating a neoplastic disease comprising
`administering to a patient in need thereof a combination of a
`VEGF inhibitor with irinotecan.
`2. The method according to claim 1, wherein the VEGF
`inhibitor is a VEGF Trap.
`3. The combination according to claim 2, wherein the
`amount of VEGF Trap represents from 10% to 80% by weight
`of the combination.
`4. The method according to claim 1, wherein said method
`does not include any other chemotoxic derivative having a
`therapeutically synergistic effect in the treatment of neoplas
`tic diseases.
`5. A product comprising a VEGF inhibitor and irinotecan,
`as a combined preparation for simultaneous, separate or
`sequential use in anticancer therapy.
`
`6. The product according to claim 5, wherein the VEGF
`inhibitor is a VEGF Trap.
`7. The product according to claim 6, wherein the amount of
`VEGF Trap represents from 10% to 80% by weight of the
`combined weight of VEGF Trap and irinotecan.
`8. The product according to claim 5 which does not include
`any other chemotoxic derivative having atherapeutically Syn
`ergistic effect in the treatment of neoplastic diseases.
`9. A combination containing a VEGF inhibitor with irino
`tecan.
`10. The combination according to claim 9, containing a
`VEGF Trap with irinotecan.
`11. The combination according to claim 10, containing
`from 10% to 80% by weight of VEGF Trap.
`
`c
`
`c
`
`c
`
`c
`
`c
`
`Regeneron Exhibit 2015
`Page 05 of 05
`
`