AU621046B2

AU621046B2 - Method for therapy of leukemias and certain other malignancies

Info

Publication number: AU621046B2
Application number: AU14878/88A
Authority: AU
Inventors: Mikael Jondal; Anders Rosen
Original assignee: Astra AB
Current assignee: AstraZeneca AB
Priority date: 1987-03-11
Filing date: 1988-03-10
Publication date: 1992-03-05
Anticipated expiration: 2008-03-10
Also published as: SE8701004D0; FI885177A0; WO1988006891A1; EP0305468A1; AU1487888A; FI885177A; JPH01502592A

Description

t.

AU-AI-14878/88 ~Rk~ P CT WORLD INTELLECTUAL PROPERTY ORGANIZATION INTERNATIONAL APPLICATION PU6S 2 U E EPTE OOPERATION TREATY (PCT) (51) International Patent Classification 4 (11) International Publication Number: WO 88/ 06891 A61K 39/00, 37/02, 35/12 Al (43) International Publication Date: 22 September 1988 (22.09.88) (21) International Application Number: (22) International Filing Date: 10 PCT/SE88/00118 March 1988 (10.03.88) (31) Priority Application Number: (32) Priority Date: (33) Priority Country: 8701004-7 11 March 1987 (11.03.87)

SE

(71) Applicant (for all designated States except US): AK- TIEBOLAGET ASTRA [SE/SE]; S-151 85 Sbdertalje

(SE).

(72) Inventors; and Inventors/Applicants (for US only) JONDAL, Mikael [SE/SE]; Strandvigen 53, S-115 23 Stockholm (SE).

ROSEN, Anders [SE/SE]; Gustafsvagen 17, S-171 49 Solna (SE).

(74) Agents: MIKSCHE, Gerhard et al.; AB Astra, Patent and Trademark Department, S-151 85 S6dertilje (SE).

(81) Designated States: AT (European patent), AU, BE (European patent), CH (European patent), DE (European patent), DK, FI, FR (European patent), GB (European patent), IT (European patent), JP, LU (European patent), NL (European patent), NO, SE (European patent), US.

Published With international search report.

A.O.J.P. 17 NO V 1988

AUSTRALIAN

OCT 1988 PATENT OFFICE (54) Title: METHOD FOR THERAPY OF LEUKEMIAS AND CERTAIN OTHER MALIGNANCIES (57) Abstract A method for the treatment of malignantly transformed cells in animals and man, which comprises the administration of a therapeutically adequate amount of specified B-cell growth and differentiation factors.

i i i; t u

'I

11 ii: ii .:il i; Field of the invention The present invention relates to a novel pharmaceutical composition suitable for the treatment of B-cell leukemias, especially B-cell chronic lymphocyte leukemia, more particular as hereinafter indicated.

The use of B-cell growth factors, and antibodies that mimic these, can be used for the induction of differentiation in certain malignant disorders. We describe here the use of certain factors, and antibodies, for the purpose indicated.

General outline of the invention and introduction Cancer cells are characterized by uncontrolled growth. For some time there has been a concept that growth can be suppressed by inducing these cells to differentiate into a •ego non-proliferative state. Clinical trials have also been done in different leukemias with differentiation-inducing agents such as vitamins and interferons. However, no such trials have been done with more specific growth and differentiation factors, or antibodies, which only react with defined receptor structures. The present invention S20 uses such specific factors for cancer treatment, in 4* combination with supporting, non-specific agents, as •hereinafter defined.

The development of normal cells into cancer cells is a i multi-step process. During malignant transformation some cell types, for example some B lymphocytes (reference i), acquire the ability to express receptors for defined growth factors and respond to these by-proliferation or maturation. Receptor expression is one important step in the progression towards a fully malignant phenotype. The tumor cells are thus "frozen" at a specific differentiation 1 stage. This block is, however, not irreversible. We here e present pharmaceutical compositions containing specified iS B-cell growth factors, and monoclonal antibodies binding to 2 corresponding receptor structures, to be used in combination with co-factors, for the induction of terminally differentiated cells (end cells) which do not further divide. The growth factors and co-factors are described. The strategy of clinical treatment is exemplified with B-cell chronic lymphocytic leukemias (B-CLL), which were induced to further differentiation (to a more mature stage) signified by impaired capacity to proliferate and the expression ofa plasmacytoid morphology, as judged by surface markers, cytoplasmic immunoglobulin, and endoplasinatic reticulum.

To grasp the concept of differentiation therapy it is important to understand how normal cells develop. In the bone marrow, different functionally specialized cell types develop as a result of differentiation (committment) of the multipotent stem cells. This differentiation gives rise to precursors of various cell linages (B-cell linage, T-cell linage, myeloid linage). Subsequent phenotypic changes of such unipotent cells into end cells is called maturation or terminal differentiation.

The activation of human B-cells from a resting stage, leading into further differentiation and maturation and the terminal stage proceeds 0 through at least two steps.

1) The triggering step, where the cells are exposed to activating factors, so-called competence inducing agents. For the B-cell series these are: Antigens; anti-immunoglobulins (anti-idiotypes); interleukin 5 1, 2 and 3 and sub-components thereof, interleukin 4 (IL4) and 0• antibodies to the IL4-receptor; reagents acting on the C3d-receptor S. (CDllc), such as polymerized complement 3d or antibodies to the C3d receptor (anti-gpl40); anti-gp35 (CD20). Phorbol esters, such as TPA or PHA are used experimentally in vitro as potent competence-inducing agents, but these can however only serve as models since they are toxic and incompatible with clinical use. The phorbol esters act on protein kinase C (PKC) and in their function mimic biologically active agents.

Other experimental competence-inducing ace-ts of importance are: solid chase protein-A; inactivated Staphylcc:::..~ A,reus Cowan I (SAC); Poke weed Mitogen (PWM); non-transforming or inactivated Epstein-Barr Virus (EBV) (from the non-transforming strain P3HRI or UV-inactivated virus) lipopolysaccharides (LPS).

i 1 1 ij j 'WO 88/06891 PCT/SE88/00118 2) The progression step. The triggering step induces receptors for various progression signals such as IL-2; B-cell'growth factor II or TRF, now called IL5; low molecular weight BCGF (12K BCGF); Namalwaderived 60K BCGF; antibodies to CD23 (a p4 5 protein expressed on the B-cell surface of IgM+, IgD+ cells, and a potential receptor for 12K BCGF); antibodies to CDw40, a p50 antigen present on B-cells and on urinary bladder carcinoma cells, but also on cervical and lung carcinoma cells (reference furthermore BSF2 (previously called B-cell differentiation factor (BCDF).

The following list is a brief explanation of abbreviations used in the present specification.

BCDF: B-cell differentiation factor BCGF: B-cell growth factor B-CLL: B-cell chronic lyrphocytic leukemia BSF: B-cell stimulating factor C3d: Sub-component of complement factor C3 CD23: A p45 protein expressed on cells of the B-lymphocyte linage, particularly IgM and IgD positive cells A p50 protein expressed on B-cells and on bladder carcinoma cells EBV: Epstein-Barr virus Glycoprotein 35K molecular weight, belonging to the CD20 group (cluster of differentiation group) gpl40: Glycoprotein 140K molecular weight, with C3d-receptor function IgD: Immunoglobulin class D IgM: Immunoglobulin class M IL-1, IL-2, IL-3, IL-4, IL-5: Interleukin 1, 2, 3, 4, LPS: Lipopolysaccharides Molt4: A T-lymphoma derived cell line p4 5 A 45K molecular weight membrane protein PMA: 4-phorbol 12-myristate 13-acetate PWM: Poke weed mitogen SAC: Staphylococcus aureus Cowan I Solid phase protein-A: Matrix (Sepharose for example) -bound protein-A TPA: Tumor promoting agent TRF: T-cell replacing factor 14 1 O4 T-T hybridoma: A somatic cell hybrid between two different T-cells Detailed description of the invention The present invention provides a novel pharmaceutical composition suitable for the treatment of B-cell leukemias, especially B-cell chronic lymphocytic leukemia, comprising a synergistic combination of: the B-cell growth and differentiation factor BCGF 12K, 10 obtainable from T-T-hybridoma MP6;and e a co-factor consisting of interleukin 2, capable of inducing malignantly transformed cells to express receptors for said factor In practice, the pharmaceutical compositions of the invention may be formulated, if desired, so as to be suitable for administration by infusion. Alternatively, if desired, the pharmaceutical compositions of the invention may be formulated so as to be suitable for administration S by intramuscular deposition.

20 The BCGF 12K factor utilized in the pharmaceutical compositions of the invention is further described in the specification on page 7 at lines 9-22. A BCGF 12K factor can be obtained from the T-T-hybridoma MP6. The synergistic effect obtainable by the combination of the said BCGF 12K and IL-2 is to be found in Table 3 hereinafter, by a comparison of the following figures: BCGF 12K factor alone: 161 co-factor IL-2 alone: 256 combination of BCGF 12K and IL-2: 5 998 These data clearly establish an unobvious synergistic effect.

Thus, the pharmaceutical compositions of the invention are suitable for the administration of therapeutically adequate amounts of the relevant growth and differentiation factors so identified, said components being included within those listed in Tables 1 and 2 below.

The relevant growth and differentiation factors referred to above are included within those listed in Table 2 and Table 1 and are substances which are known as such. They are, however, not in every instance known to have therapeutic utility.

In clinical practice, the relevant growth and differentiation factors are administered in a manner which is analogous with known ways of administering medicaments for the treatment of cancer. Thus, administration will preferably be made by infusion or by intramuscular deposition.

eeeee: The amount in which the relevant growth and different- •iation factors are administered will vary within a wide :range and will depend on various circumstances such as the 20 severity of the disease and the age and the state of the patient. As an example of a suitable dosage interval can be mentioned from 10 000 to 300 000 Units of growth and differentiation factor per kg bodyweight per 24 hours. An amount of 200 000 U per kg bodyweight per 24 hours will sometimes be adequate.

The following general description is provided for the purpose of illustrating the pharmaceutical compositions containing the relevant growth and differentiation factors according to the invention. Table 1 lists inter alia the co-factors used therein. The designation E indicates that the co-factor mainly is experimental and has possible use for diagnostic purposes. The designation C indicates that the co-factor has clinical use.

.T

~a 6 Table 1. Co-factors (Competence inducing agents) .r E Phorabol esters such as TPA E Antigens C Anti-Immunoglobulins (anti-idiotypes) C Interleukin 1 and sub-components thereof C Interleukin 2 and sub-components thereof C Interleukin 3 and sub-components thereof C Interleukin 4 (BSF1) C Anti-IL4-receptor antibodies E Poke weed mitogen E Lipopolysaccharides E Epstein Barr virus, non-transforming or inactivated C C3d receptor (CDllc) reactive agents C' and antireceptor (gp 140) antibodies C Anti-gp35 E SAC, Inactivated Staphylococcus aureus Cowan I E Solid-phase protein A C Interferons (alfa, beta and gamma) C Vitamins, in particular vitamin A, D, and biologically active derivatives The following Table 2 lists inter alia B-cell growth and differentiation factor used in the pharmaceutical compositions according to the invention. All listed factors have clinical use.

25 Table 2. B-cell Growth and Differentiation Factors (progression signals) BCGF 12K and high molecular weight BCGF derived from Thelper cells and T-T hybridomas BCGF 30K (cleavage product of the CD23) derived from B-cell lines BCGF 60K derived from the Namalwa cell line BSF2 also called BCDF Interleukin 5, previously called BCGF II or TRF Anti-CD23 antibodies A Ij- WOg 88/06891 PCT/SE8800118 7 (p50) antibodies, present on B-cells and bladder carcinoma cells Anti-BCGF receptor antibodies Gamma-interferon Interleukin 2 and sub-components thereof Definition of the B-cell growth factors The producer cells of the BCGF's can be T-helper cells immortalized by somatic cell hybridization with a T-lymphoma called Molt4. One of these BCGF-producer cell lines (T-T-hybridoma MP6) are described in detail in reference 3.

The biochemical properties of this BCGF are: 1) Ammonium sulphate precipitable at 80-90% saturation, O°C 2) Heat labile at 56 0 C, 30 min.

3) Protease sensitive 4) Glycoprotein nature, with binding capacity to mannose-specific lectins HPLC-gelfiltration analysis gives an apparent molecular weight of 12-14K, as well as high molecular weight component Biological properties The growth and differentiation factors according to Table 2 stimulate the proliferation and differentiation of normal and B-CLL lymphocytes pretreated or co-treated with co-factors as shown in Table 1 (anti-IgM, SAC, TPA).

Target cells in clinical situations: Target cells in clinical situations are all such malignantly transformed cells that express receptors for factors according to Table 2 and respond to these factors by differentiation, including all those malignant cells that can be induced to express receptors for the factors described in Table 2 and respond to these. Such induction can be exerted by the co-factors described in Table 1 or by other means.

i I _1 WO 88/06891 PCT/SE88/00118 Table 3. Response of B-CLL leukemic cells to a B-cell growth factor, derived from T-T hybridoma MP6, in combination with co-factors IL1 and IL2 B-cell growth factor Co-factor Response of B-CLL leukemic cells cpm SI MP6 241 below 1 IL-1 161 below 1 IL-2 256 below 1 IL-1 and IL-2 373 below 1 MP6 IL-1 456 below 1 MP6 IL-2 5 988 11.6 MP6 IL-1 and IL-2 28 550 55.5 Medium control 514 Legend: Isolated B-CLL leukemic cells reference 3. The response was H-thymidine at 72 h.

were stimulated as described in measured in incorporation of SI stimulation index, experimental value divided by back-ground control.

i;

Claims

1. A pharmaceutical composition suitable for the treatment of B-cell leukemias, especially B-cell chronic lymphocytic leukemia, comprising a synergistic combination of: the B-cell growth and differentiation factor BCGF 12K, obtainable from T-T-hybridoma MP6; and a co-factor consisting of interleukin 2, capable of inducing malignantly transformed cells to express 0 receptors for said factor

2. A pharmaceutical composition according to claim 1, formulated so as to be suitable for administration by infusion.

3. A pharmaceutical composition according to claim 15 1, formulated so as to be suitable for administration by Sintramuscular deposition.

4. A pharmaceutical composition according to any one of claims 1 to 3, formulated to deliver a dosage interval from 10 000 to 300 000 Units of growth and differentiation factor per kg bodyweight per 24 hours. Sca

5. A pharmaceutical composition according to any one of claims 1 to 3, formulated to deliver a dosage interval of 200 000 Units of growth and differentiation factor per kg bodyweight per 24 hours. DATED this 16th day of December, 1991 AKTIEBOLAGET ASTRA, By its Patent Attorneys, E. F. WELLINGTON CO., i S. Wellingtpt)