CA2018133A1 - Use of xylan polyhydrogensulfates for the therapy of disorders based on cell proliferation - Google Patents
Use of xylan polyhydrogensulfates for the therapy of disorders based on cell proliferationInfo
- Publication number
- CA2018133A1 CA2018133A1 CA002018133A CA2018133A CA2018133A1 CA 2018133 A1 CA2018133 A1 CA 2018133A1 CA 002018133 A CA002018133 A CA 002018133A CA 2018133 A CA2018133 A CA 2018133A CA 2018133 A1 CA2018133 A1 CA 2018133A1
- Authority
- CA
- Canada
- Prior art keywords
- carcinoma
- dalton
- compound
- treatment
- formula
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
BEHRINGWERKE AKTIENGESELLSCHAFT 89/B 024 - Ma 730 Dr. Ha/Sd Abstract of the disclosure The use of xylan polyhydrogensulfates for the therapy of disorders based on cell proliferation The use of compounds of the formula I
I
in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixtures with a mean molecular weight of about 1,000 to 20,000 Dalton, preferably of about 5,000 to 12,000 Dalton, especially about 6,000 Dalton (? = 9), or of one of the pharmacologically acceptable acid addition salts thereof, for the prepara-tion of a pharmaceutical for the therapy of disorders based on uncontrolled and undifferentiated cell growth, is described.
I
in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixtures with a mean molecular weight of about 1,000 to 20,000 Dalton, preferably of about 5,000 to 12,000 Dalton, especially about 6,000 Dalton (? = 9), or of one of the pharmacologically acceptable acid addition salts thereof, for the prepara-tion of a pharmaceutical for the therapy of disorders based on uncontrolled and undifferentiated cell growth, is described.
Description
z;~
B~ERINGhERR~ ARTI~NOE S~L~SC~a~T 89/B 024 - Na 730 Dr. ~a/Sd The ~e of ~ylan polyhydrogensulfates for ~he therapy of di~orders ba~ed on ~ell proliferation _ The in~ention relates to the use of xylan p~lyhydrogen-sulfates for the preparation of a pharmaceutical for the therapy of disorders basQd on uncontrolled and undif-ferentiated cell growth.
Xylan polyhydrogensulfates are disclosed in DE-~36 01 136. The following formula I is given therein for these compounds;
H~
: /O~H/
H OSO~N- _ n An anti-retroviral action i~ al~o de~cribed for these compounds therein. It has furthermore been dis~losed that xylan polyhydrogensulfate inhibits ~he binding of bFGF
(basic fibroblast growth fac~or) to an adrenocortical carcinoma cell line (SW 13) (A. Wellstein et al., Proc.
of the American Association for Cancer Research (1989) 30, 583).
It has now been found/ surprisingly, that compounds of the formula I inhibit oncogene-encoded Xinases and growth factor receptor tyrosine kinases. Hence they are suitable for controlling diseases in which growth factor receptor~
play a part, especially p60riasis and onco~es.
~he expression of oncogenes in a mammalian cell i8 associated with the transition from the normal to the transformed type of cell, which then becomes a cancer cell. The transformation can be induced by infection of a cell with a retro~irus. A well-known e~ample i8 RoU8 sarcoma virus infection of chickens, which subsequently develop cancer. The relevant oncogene which is responsible for the malignant transformation has been called the "SRC" (sarcoma) gene. Many of the oncogenes now known are characterized by the expression of a protein with kinase activi~y. The anzymes catalyze the transfer of the terminal phosphate group of ATP to an amino acid. In contrast to many other protein kina~es which transfer the phosphate group ~o a seryl or threonyl radical, most of the oncogene-encoded kinases phosphory-late a tyrosyl radical of the protein chain. Apart from this, it is known that products of oncogenes, namely those of the v-mos, v-mil and v-raf oncogenes, have serine/threonine-specific protein kinase activity.
Tyrosine kinase activity is also expressed in growth factor receptors; recent results have now shown that various diseases in ~hich the proliferation of cell~
plays a part, for example psoriasis~ and the growth of many tumors depend on the presence of growth factors, such as epidermal growth factor (EGF), transforming growth factor alpha (TGF alpha), platelet derived growth factor (PDGF) or fibroblas~ growth factor ~basic FGF, acidic FGF). Binding of the growth factor to its receptor is followed by stimulation of tyrosine kinase which is an intrinsic component of the growth factor receptor.
~his is why an inhibitor of the tyrosine kinase of a growth factor receptor inhibits cell growth and thus, for example, also tumor growth and the ~preading of tumors.
It can therefore be employed in tumor therapy and in the therapy of all diseases in which cell growth plays a part (for example psoriasis~.
~he invention therefore relates to the use of a compound of the formula I
B~ERINGhERR~ ARTI~NOE S~L~SC~a~T 89/B 024 - Na 730 Dr. ~a/Sd The ~e of ~ylan polyhydrogensulfates for ~he therapy of di~orders ba~ed on ~ell proliferation _ The in~ention relates to the use of xylan p~lyhydrogen-sulfates for the preparation of a pharmaceutical for the therapy of disorders basQd on uncontrolled and undif-ferentiated cell growth.
Xylan polyhydrogensulfates are disclosed in DE-~36 01 136. The following formula I is given therein for these compounds;
H~
: /O~H/
H OSO~N- _ n An anti-retroviral action i~ al~o de~cribed for these compounds therein. It has furthermore been dis~losed that xylan polyhydrogensulfate inhibits ~he binding of bFGF
(basic fibroblast growth fac~or) to an adrenocortical carcinoma cell line (SW 13) (A. Wellstein et al., Proc.
of the American Association for Cancer Research (1989) 30, 583).
It has now been found/ surprisingly, that compounds of the formula I inhibit oncogene-encoded Xinases and growth factor receptor tyrosine kinases. Hence they are suitable for controlling diseases in which growth factor receptor~
play a part, especially p60riasis and onco~es.
~he expression of oncogenes in a mammalian cell i8 associated with the transition from the normal to the transformed type of cell, which then becomes a cancer cell. The transformation can be induced by infection of a cell with a retro~irus. A well-known e~ample i8 RoU8 sarcoma virus infection of chickens, which subsequently develop cancer. The relevant oncogene which is responsible for the malignant transformation has been called the "SRC" (sarcoma) gene. Many of the oncogenes now known are characterized by the expression of a protein with kinase activi~y. The anzymes catalyze the transfer of the terminal phosphate group of ATP to an amino acid. In contrast to many other protein kina~es which transfer the phosphate group ~o a seryl or threonyl radical, most of the oncogene-encoded kinases phosphory-late a tyrosyl radical of the protein chain. Apart from this, it is known that products of oncogenes, namely those of the v-mos, v-mil and v-raf oncogenes, have serine/threonine-specific protein kinase activity.
Tyrosine kinase activity is also expressed in growth factor receptors; recent results have now shown that various diseases in ~hich the proliferation of cell~
plays a part, for example psoriasis~ and the growth of many tumors depend on the presence of growth factors, such as epidermal growth factor (EGF), transforming growth factor alpha (TGF alpha), platelet derived growth factor (PDGF) or fibroblas~ growth factor ~basic FGF, acidic FGF). Binding of the growth factor to its receptor is followed by stimulation of tyrosine kinase which is an intrinsic component of the growth factor receptor.
~his is why an inhibitor of the tyrosine kinase of a growth factor receptor inhibits cell growth and thus, for example, also tumor growth and the ~preading of tumors.
It can therefore be employed in tumor therapy and in the therapy of all diseases in which cell growth plays a part (for example psoriasis~.
~he invention therefore relates to the use of a compound of the formula I
2~
- 3 ~
~H ~1 I
/o~/ CSO,N~
_ ' _ n in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixture~ with a mean molecular weight o~ about 1,000 to 20,000 Dalton~ preferably of about 5,000 to 12tO00 Dalton, especially about 6,000 Dalton (n = 9), for the preparation of a pharmaceutical for inhibiting oncogene-encoded tyro~ine kinases, growth factor receptor ~yrosine kinases and growth factor/-receptor interaction and thus for controlling disorders based on cell proliferation, preferably psoriasis, and oncoses.
Tumors of this type are, in particular, carcinomas and sarcomas such as renal carcinoma, mammary carcinoma, prostate carcinoma, pulmonary carcinoma, ovarian car-cinoma or rhabdomyosarcoma or melanoma.
The compounds according to the invention have advan-tageous pharmacological properties in that they inhibit the growth and the spread of tumors and can therefore be used in tumor therapy and in the therapy of all disorders based on cell proliferation, that is to say disorders which are characterized by uncontrolled and undifferen-tiated cell growth.
For this purpose it is possible to use the compounds of the formula I themselves also in combination or their pharmacologically acceptable acid addition sal~s.
A suitable pharmaceutical is prepared by converting the active substances, where appropriate wi~h auxiliaries and/or excipients, into a sui~able dosage form.
The examples which follow explain the in~ention.
Examples Assay for inhibi~ion of EG~ receptor tyrosine ki~ase The star~ing material for the tyrosine kinase activity was the human tumor cell line A 431 (ATCC CRL 1555), which was cultivated in ~PMI 1640 medium ~ 10% FCS. This cell line expresses on the cell surface a large number of EGF receptors which have tyrosine kinase acti~ity.
The cells were cultivated almo~t to confluence, washed with PBS (phosphate-buffered saline, p~ 7.2), scraped off the culture flask and incubated at 4C for 1 h, treated 20x in a Potter and centrifuged at lOOOxg for 30'. The supernatant was centrifuged at 20,000xg for a further 20 min, and the pellet was taken up in 100 ~1 per 1 x 106 cells as membrane preparation.
The tyrosine kinase activity of the EGF receptor was measured with poly(Glu, Ala, Tyr 6:391) as substrate. The cell membranes were treated with 1000 nM EGF at RT for 15' and then added to the mixture which contains the inhibitor, substrate (3 mg/ml), Mg2'/Mh~ (8 mM/ 1.6 mM)~
0.16% Triton X-100 and sodium ortho~anadate ~20 ~M) in ~ 100 mM HEPES (N-2-hydroxyethyl-piperazine-N~-2-ethanesul-fonic acid), pH 7.5, and preincubated, and the reaction was started by adding gamma~32P-ATP (32 ~M). After 15~ at 30C, the substrate was precipitated with 10% TCA (tri-chloroacetic acid), filtered on a hillititer fil~ration plate (Millipore Corporation, Mass., US~), washéd and dried. The incorporation of 32p was determined using a 1 iquid scintillation counter.
Results:
23~ 3 ~ 5 -Xylan polyhydrogensulfate (n = 9) ~as tested at a maximum concentration of 1460 ~g/ml and subjected to stepwise 1:10 dilution. The IC50 was defined as the concentration at which 50% of ~he initial enzyme ac~ivity was inhibited. A fiyure of 6 ~g/ml was determined for EGF
receptor tyrosine kinase (Table 1).
Table 1 Inhibition of the enzymatic activity of protein kinases-by xylan polyhydrogensulfate (formula I in which n = 9) IC50 in ~g/ml ICsOin~g/ml EGF receptor cAMP-depen-kyrosine dent protein kinase kinase _ Xylan polyhydrogensulfa$e (n=9) 6 884 .
Assay for inhibition of 3', 5'-cAMP~dependent protein kinase The catalytic subunit of cAMP-dependent protein kinase (Sigma) was reconstituted as described by Sigma (Sigma Chemical Co., St. Louis, MO, USA). The enzyme activity was measured using kemptide (Sigma) (Leu-Arg-Arg-Ala-Ser-Leu-Gly) as substrate. The inhibitor was preincubated with enzyme, substrate (190~ M), Mg2~ (5 ~M), 0.25 mg/ml BSA and 3.75 mM mercaptoethanol in 50 mM MOPS (4-morpho-linopropanesulfonic acid), pH 6.9. The reaction was started by adding gamma-32P-ATP (40 ~ N). After 15' at 30C, an aliquot was applied to p81 ion exchanger (2 x 9 2 cm; Whatman Paper Ltd, Great Britain), dipped i~ 75 mM
H3PO4, washed and dried, and $he incorporation of 32p was determined using a liquid scintillation counter.
21J~131 33 Results:
Xylan polyhydrogensulfate (n=9) was tested at a ma~imum concentration of 1142 ~g/ml and sub~ected to ~tepwise 1:10 dilution. The IC50 was de~ined a~ the concentration at which 50~ of the ini~ial enzyme activity was inhibited. The IC50 for cAMP-dependent protein kinas~ was 884 ~g/ml (Table 1~, i.e. xylan polyhydrogensulfate (n=9) is a specific inhibitor of tyrosine kinase.
Inhibition of non-adherent growth of A 549 (human pul-monary carcinoma cell line~, A 204 (human rhabdomyosar-coma cell line), Ovar Cal (human ovarian carcinoma cell line), MDA-MB231, MDA-MB468 (human mammary carcinoma cell lines), ~nCaP (human prostate carcinoma cell line) and NRK (normal rat kidney fibroblasts).
The assay was carried out by the method de~cribed by Hamburger and Salmon (J. Natl. Cancer Inst. 6S, 981-988, 1981) with the modifications described below.
Conditioned medium was replaced by RPMI 1640 Medium (A
549, A 204, Ovar Cal) or IMEM (all other cell lines) +
10% FCS (fetal calf serum). As a consequence of the high cloning rate of the tumor cell lines in soft agar, the number of tumor cells per plate was reduced to 3 x 103 for A 549 and to 10 x 103 for all the others.
The cells were plated out as the upper layer in a two-layer agar sy~tem in accordance with the Hamburger and Salmon method (J. ~atl. Cancer Inst. 66, 981-988, 1981), with various concentrations of the test substance +
additional EGF (epidermal growth factor) (1 nM3 or additional bFGF (basic fibroblast growth factor) (100 ng/ml) being mixed with the upper agar layer before the cells were plated out.
The plates were incubated in an incubator with 5% CO2, 20~ 2 and 95~ relative humidity for 10 days ~A 549 and A
;2~?1~
204), 18 ~ays (Ovar Cal) or 1~ days (all other cells).
After this tLme, colonies with a diameter greater than 60 ~m were counted using an Lmage analyzer. The IC50 has been reported as the concentration of substance at which the number of colonies was reduced to one half.
The coefficient of variation on repetition of the experi-ments was less than 15~.
Result:
Xylan polyhydrogensulfate (n=9) inhibits the prolifera-tion of cell lines which have receptors, containingtyrosine phosphokinase, for various growth factors. This is why cell growth is inhibi~ed irrespective of whether the particular growth factor was also added or not (see 1, 2 and 4 in Table 2).
There is likewise inhibition in the case of cells which grow only in the presence of a growth factor (in the cell culture) (see 3). Cells not dependent on growth factors (see 5) are unaffected.
The IC50 of xylan polyhydrogensulfate (n=9) for continuous incubation was determined from the dose-effect plot. The results were:
Table 2 Tumor cell line IC50 (~g/~l) 1. A 549 29 (- E~F) S 59 (~ EGF~
2. Ovar Cal 4 (- bFGF) 6 (~ bFGF) 3. A 204 34 (+ b~F) `
(without bFGF no colonies) 4. LNcap 200 (~ bFGF) NRK ~GF-dependent 60 5. MDA 231 Inhibition of human tumor growth in the renal capsule assay (nude mouse) 1. Preparation of the tumor The human tumors to be tested are routinely maintained and passaged in nu/nu CDl mice~ The tumor is removed under sterile conditions, connective tissue and necro~ic portions are removed, and the tumor is comminuted and disintegrated with 20 ml of the following solution (100 ml of PBS without Ca2+ and Mg2~ with 800 mg of cul-lagenase (Serva 0.6-0.8 U/ml) + 2 mg of DNA~e). Incuba-~ion: 2 h, 37C with shaking. The cell ~uspension is then filtered through a nylon net (pore diameter 51 ~m) and washed three times with Ca2+- and Mg~-free PBS. The cells are then counted and pelleted.
20~ ~ 3~
g 2. Embedding the cells in fibrin pieces The interior wall of a 50 ~1 glass capillary ~diameter 1.5 mm) is wetted with a thrombin/CaCl2 ~olu~ion (50 unit of thrombin in 1 ml of 40 mmol/l CaClz).
~he tumor cell pellet is suspended in a coagulable fibrinogen solution~ such as RBeriplast (Behringwerke AG), and diluted 1:2 with RPNI ~ 15% FCS to a final c~ll concentration of 107 cell~/50 ~1. This ~uspension is rapidly a~pirated into the qlass capillarie~.
After the cell-fibrin mixture has Rolidified (about 5 min at room temperature) the fibrin adhesive is forced with compressed air out of the glass capillaries into Petri dishes. The fibrin adhesive is cut up into 2 mm pieces (corresponding to 5 ~ 105 cells/piece), and the pieces are stored in RPMI + 15~ FCS until implanted.
3. Implantation The animals are anesthetized with Nembutal (diluted 1:7 with physiological saline; 0.1 ml/10 g of mouse). The kidney is exposed by a 1.5 cm-long incision into the flank. The renal capsule is cut open and a single fibrin piece is transferred under the renal capsule. Then the two diameters of the implanted piece are measured using a microscope with an ocular micrometer. Th~ t~mor size is determined from V = a x b, where V = tumor size, a =
largest diameter of tumor and b = tumor diameter at right angles to a.
The peritoneum is then closed with tissue adhesive (Histoacryl~, and the skin is ~lipped.
Treatment The substance is administered i.v. each day on days 2-10 with the maximum tolerable dose and with 2/3 of the L3~
maxLmum tolerable dose found in preliminary tests.
Measurement On day 20 after the implantation, the animals are sacri-ficed, the kidney is exposed and the tumor size iS again measured. The efficacy of the ~est substance is deter-mined by the inhibition of tumor growth.
The relative tumor size VR is calculated using YR = Vt/VO~
where ~t is the tumor size at the end of the experiment (day 20) and V0 is the size of the tumor fibrin piece on the day of implantation.
Then the median relative tumor size in the treated group (VT) is related to the corresponding median relative tumor si~e of the controls (VC)~ snd T/C % = VT~VC X 100 i8 calculated.
~he statistical significance of the antitumor effect i8 determined usin~ the Wilcoxon U test. ~he relati~e tumor si~es in the treated group are compared with the cor-responding relative si~es in ~he control group, and the change in the tumor size is regarded as substance-specific only if it is statistically significant with pless than 0.05.
~J~ 3~
Result:
Table 3 Tumor Dose Activity (T/C %) .
LXF 529 20 mg/kg 70 (human bronchial carcinoma) 30 mg/kg 71 MCF7 20 mg/kg 66 10~human mamMary carcinoma) 30 mg/kg 65 n.s.
Ovar-6 20 mg/kg 78 n.s.
(human ovarian carcinoma) 30 mg/kg 66 PaTu 8902 20 mg/kg 56 (human pancreatic carcinoma) 30 mg/kg 27 ~T 29 20 mg/kg 71 (human colon carcinoma) 30 mg/kg 76 n.s.
n.s. = not significan~
Inhibition of human tumor growth in subcut~neously growing human t~mors (nude mou~e) The tested human tumors are routinely maintained and passaged in nu/nu CD~ mice. ~he $umors are tested at each S third passage for their human charactPr by immunohist-ology using monoclonal an~ibodies. ~he ~umor is remo~ed under s~erile conditions and cut into small pieces of volume about 1~10 mm3. One tumor piece is implanted subcutaneously in the side of each nude mouse. Af~er 7-14 days the tumor piece has grown into the surroundingtissue, and the ~umor size is determined from the 2 diameters using the following fonmula: V = a x b (see above).
This procedure is repeated twice a week, and only animals 1~ with progressive tumor growth are randomi~ed to the control group and the group to be ~rea~ed. Five mice are used per group. Starting on the day of randomization the animals are ~reated intravenously in accordance with ~he treatment regimen which is indicated in the results section. Two different dosages are used for each sub-stance. The selected dosages are identical to the maximum tolerable dose (MTD) which was determined beforehand for each test substance in an extra experiment with nude mice.
The two tumor diameters are measured for each mouse twice a week, and the individual tumor area is calculated using the abovementioned formula.
Evaluation:
The relative tumor size VR and T/C ~ are calculated as above.
The statistical significance of the antitumor effect is determined using the Wilcoxon U test. The relative tumor sizes in the treated group are compared with the cor-3~:~
responding relative sizes in the control group on the same day of the experiment. The change in the tumor area is regarded as substance-speci~ic only if it is statisti-cally significant with p less than O . 05 .
Result:
Table 4 5~e Tm~r Dose Reyime Acti~ity (day 9);
(T/C %) .
Xylan poly- B 17 15 mg~bg QD9 x i.v. 61.4 ~- (~aan sulfate bron~
(n=9) camL%~) 20 mg/hg QD9 x i.v. 68.
~H ~1 I
/o~/ CSO,N~
_ ' _ n in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixture~ with a mean molecular weight o~ about 1,000 to 20,000 Dalton~ preferably of about 5,000 to 12tO00 Dalton, especially about 6,000 Dalton (n = 9), for the preparation of a pharmaceutical for inhibiting oncogene-encoded tyro~ine kinases, growth factor receptor ~yrosine kinases and growth factor/-receptor interaction and thus for controlling disorders based on cell proliferation, preferably psoriasis, and oncoses.
Tumors of this type are, in particular, carcinomas and sarcomas such as renal carcinoma, mammary carcinoma, prostate carcinoma, pulmonary carcinoma, ovarian car-cinoma or rhabdomyosarcoma or melanoma.
The compounds according to the invention have advan-tageous pharmacological properties in that they inhibit the growth and the spread of tumors and can therefore be used in tumor therapy and in the therapy of all disorders based on cell proliferation, that is to say disorders which are characterized by uncontrolled and undifferen-tiated cell growth.
For this purpose it is possible to use the compounds of the formula I themselves also in combination or their pharmacologically acceptable acid addition sal~s.
A suitable pharmaceutical is prepared by converting the active substances, where appropriate wi~h auxiliaries and/or excipients, into a sui~able dosage form.
The examples which follow explain the in~ention.
Examples Assay for inhibi~ion of EG~ receptor tyrosine ki~ase The star~ing material for the tyrosine kinase activity was the human tumor cell line A 431 (ATCC CRL 1555), which was cultivated in ~PMI 1640 medium ~ 10% FCS. This cell line expresses on the cell surface a large number of EGF receptors which have tyrosine kinase acti~ity.
The cells were cultivated almo~t to confluence, washed with PBS (phosphate-buffered saline, p~ 7.2), scraped off the culture flask and incubated at 4C for 1 h, treated 20x in a Potter and centrifuged at lOOOxg for 30'. The supernatant was centrifuged at 20,000xg for a further 20 min, and the pellet was taken up in 100 ~1 per 1 x 106 cells as membrane preparation.
The tyrosine kinase activity of the EGF receptor was measured with poly(Glu, Ala, Tyr 6:391) as substrate. The cell membranes were treated with 1000 nM EGF at RT for 15' and then added to the mixture which contains the inhibitor, substrate (3 mg/ml), Mg2'/Mh~ (8 mM/ 1.6 mM)~
0.16% Triton X-100 and sodium ortho~anadate ~20 ~M) in ~ 100 mM HEPES (N-2-hydroxyethyl-piperazine-N~-2-ethanesul-fonic acid), pH 7.5, and preincubated, and the reaction was started by adding gamma~32P-ATP (32 ~M). After 15~ at 30C, the substrate was precipitated with 10% TCA (tri-chloroacetic acid), filtered on a hillititer fil~ration plate (Millipore Corporation, Mass., US~), washéd and dried. The incorporation of 32p was determined using a 1 iquid scintillation counter.
Results:
23~ 3 ~ 5 -Xylan polyhydrogensulfate (n = 9) ~as tested at a maximum concentration of 1460 ~g/ml and subjected to stepwise 1:10 dilution. The IC50 was defined as the concentration at which 50% of ~he initial enzyme ac~ivity was inhibited. A fiyure of 6 ~g/ml was determined for EGF
receptor tyrosine kinase (Table 1).
Table 1 Inhibition of the enzymatic activity of protein kinases-by xylan polyhydrogensulfate (formula I in which n = 9) IC50 in ~g/ml ICsOin~g/ml EGF receptor cAMP-depen-kyrosine dent protein kinase kinase _ Xylan polyhydrogensulfa$e (n=9) 6 884 .
Assay for inhibition of 3', 5'-cAMP~dependent protein kinase The catalytic subunit of cAMP-dependent protein kinase (Sigma) was reconstituted as described by Sigma (Sigma Chemical Co., St. Louis, MO, USA). The enzyme activity was measured using kemptide (Sigma) (Leu-Arg-Arg-Ala-Ser-Leu-Gly) as substrate. The inhibitor was preincubated with enzyme, substrate (190~ M), Mg2~ (5 ~M), 0.25 mg/ml BSA and 3.75 mM mercaptoethanol in 50 mM MOPS (4-morpho-linopropanesulfonic acid), pH 6.9. The reaction was started by adding gamma-32P-ATP (40 ~ N). After 15' at 30C, an aliquot was applied to p81 ion exchanger (2 x 9 2 cm; Whatman Paper Ltd, Great Britain), dipped i~ 75 mM
H3PO4, washed and dried, and $he incorporation of 32p was determined using a liquid scintillation counter.
21J~131 33 Results:
Xylan polyhydrogensulfate (n=9) was tested at a ma~imum concentration of 1142 ~g/ml and sub~ected to ~tepwise 1:10 dilution. The IC50 was de~ined a~ the concentration at which 50~ of the ini~ial enzyme activity was inhibited. The IC50 for cAMP-dependent protein kinas~ was 884 ~g/ml (Table 1~, i.e. xylan polyhydrogensulfate (n=9) is a specific inhibitor of tyrosine kinase.
Inhibition of non-adherent growth of A 549 (human pul-monary carcinoma cell line~, A 204 (human rhabdomyosar-coma cell line), Ovar Cal (human ovarian carcinoma cell line), MDA-MB231, MDA-MB468 (human mammary carcinoma cell lines), ~nCaP (human prostate carcinoma cell line) and NRK (normal rat kidney fibroblasts).
The assay was carried out by the method de~cribed by Hamburger and Salmon (J. Natl. Cancer Inst. 6S, 981-988, 1981) with the modifications described below.
Conditioned medium was replaced by RPMI 1640 Medium (A
549, A 204, Ovar Cal) or IMEM (all other cell lines) +
10% FCS (fetal calf serum). As a consequence of the high cloning rate of the tumor cell lines in soft agar, the number of tumor cells per plate was reduced to 3 x 103 for A 549 and to 10 x 103 for all the others.
The cells were plated out as the upper layer in a two-layer agar sy~tem in accordance with the Hamburger and Salmon method (J. ~atl. Cancer Inst. 66, 981-988, 1981), with various concentrations of the test substance +
additional EGF (epidermal growth factor) (1 nM3 or additional bFGF (basic fibroblast growth factor) (100 ng/ml) being mixed with the upper agar layer before the cells were plated out.
The plates were incubated in an incubator with 5% CO2, 20~ 2 and 95~ relative humidity for 10 days ~A 549 and A
;2~?1~
204), 18 ~ays (Ovar Cal) or 1~ days (all other cells).
After this tLme, colonies with a diameter greater than 60 ~m were counted using an Lmage analyzer. The IC50 has been reported as the concentration of substance at which the number of colonies was reduced to one half.
The coefficient of variation on repetition of the experi-ments was less than 15~.
Result:
Xylan polyhydrogensulfate (n=9) inhibits the prolifera-tion of cell lines which have receptors, containingtyrosine phosphokinase, for various growth factors. This is why cell growth is inhibi~ed irrespective of whether the particular growth factor was also added or not (see 1, 2 and 4 in Table 2).
There is likewise inhibition in the case of cells which grow only in the presence of a growth factor (in the cell culture) (see 3). Cells not dependent on growth factors (see 5) are unaffected.
The IC50 of xylan polyhydrogensulfate (n=9) for continuous incubation was determined from the dose-effect plot. The results were:
Table 2 Tumor cell line IC50 (~g/~l) 1. A 549 29 (- E~F) S 59 (~ EGF~
2. Ovar Cal 4 (- bFGF) 6 (~ bFGF) 3. A 204 34 (+ b~F) `
(without bFGF no colonies) 4. LNcap 200 (~ bFGF) NRK ~GF-dependent 60 5. MDA 231 Inhibition of human tumor growth in the renal capsule assay (nude mouse) 1. Preparation of the tumor The human tumors to be tested are routinely maintained and passaged in nu/nu CDl mice~ The tumor is removed under sterile conditions, connective tissue and necro~ic portions are removed, and the tumor is comminuted and disintegrated with 20 ml of the following solution (100 ml of PBS without Ca2+ and Mg2~ with 800 mg of cul-lagenase (Serva 0.6-0.8 U/ml) + 2 mg of DNA~e). Incuba-~ion: 2 h, 37C with shaking. The cell ~uspension is then filtered through a nylon net (pore diameter 51 ~m) and washed three times with Ca2+- and Mg~-free PBS. The cells are then counted and pelleted.
20~ ~ 3~
g 2. Embedding the cells in fibrin pieces The interior wall of a 50 ~1 glass capillary ~diameter 1.5 mm) is wetted with a thrombin/CaCl2 ~olu~ion (50 unit of thrombin in 1 ml of 40 mmol/l CaClz).
~he tumor cell pellet is suspended in a coagulable fibrinogen solution~ such as RBeriplast (Behringwerke AG), and diluted 1:2 with RPNI ~ 15% FCS to a final c~ll concentration of 107 cell~/50 ~1. This ~uspension is rapidly a~pirated into the qlass capillarie~.
After the cell-fibrin mixture has Rolidified (about 5 min at room temperature) the fibrin adhesive is forced with compressed air out of the glass capillaries into Petri dishes. The fibrin adhesive is cut up into 2 mm pieces (corresponding to 5 ~ 105 cells/piece), and the pieces are stored in RPMI + 15~ FCS until implanted.
3. Implantation The animals are anesthetized with Nembutal (diluted 1:7 with physiological saline; 0.1 ml/10 g of mouse). The kidney is exposed by a 1.5 cm-long incision into the flank. The renal capsule is cut open and a single fibrin piece is transferred under the renal capsule. Then the two diameters of the implanted piece are measured using a microscope with an ocular micrometer. Th~ t~mor size is determined from V = a x b, where V = tumor size, a =
largest diameter of tumor and b = tumor diameter at right angles to a.
The peritoneum is then closed with tissue adhesive (Histoacryl~, and the skin is ~lipped.
Treatment The substance is administered i.v. each day on days 2-10 with the maximum tolerable dose and with 2/3 of the L3~
maxLmum tolerable dose found in preliminary tests.
Measurement On day 20 after the implantation, the animals are sacri-ficed, the kidney is exposed and the tumor size iS again measured. The efficacy of the ~est substance is deter-mined by the inhibition of tumor growth.
The relative tumor size VR is calculated using YR = Vt/VO~
where ~t is the tumor size at the end of the experiment (day 20) and V0 is the size of the tumor fibrin piece on the day of implantation.
Then the median relative tumor size in the treated group (VT) is related to the corresponding median relative tumor si~e of the controls (VC)~ snd T/C % = VT~VC X 100 i8 calculated.
~he statistical significance of the antitumor effect i8 determined usin~ the Wilcoxon U test. ~he relati~e tumor si~es in the treated group are compared with the cor-responding relative si~es in ~he control group, and the change in the tumor size is regarded as substance-specific only if it is statistically significant with pless than 0.05.
~J~ 3~
Result:
Table 3 Tumor Dose Activity (T/C %) .
LXF 529 20 mg/kg 70 (human bronchial carcinoma) 30 mg/kg 71 MCF7 20 mg/kg 66 10~human mamMary carcinoma) 30 mg/kg 65 n.s.
Ovar-6 20 mg/kg 78 n.s.
(human ovarian carcinoma) 30 mg/kg 66 PaTu 8902 20 mg/kg 56 (human pancreatic carcinoma) 30 mg/kg 27 ~T 29 20 mg/kg 71 (human colon carcinoma) 30 mg/kg 76 n.s.
n.s. = not significan~
Inhibition of human tumor growth in subcut~neously growing human t~mors (nude mou~e) The tested human tumors are routinely maintained and passaged in nu/nu CD~ mice. ~he $umors are tested at each S third passage for their human charactPr by immunohist-ology using monoclonal an~ibodies. ~he ~umor is remo~ed under s~erile conditions and cut into small pieces of volume about 1~10 mm3. One tumor piece is implanted subcutaneously in the side of each nude mouse. Af~er 7-14 days the tumor piece has grown into the surroundingtissue, and the ~umor size is determined from the 2 diameters using the following fonmula: V = a x b (see above).
This procedure is repeated twice a week, and only animals 1~ with progressive tumor growth are randomi~ed to the control group and the group to be ~rea~ed. Five mice are used per group. Starting on the day of randomization the animals are ~reated intravenously in accordance with ~he treatment regimen which is indicated in the results section. Two different dosages are used for each sub-stance. The selected dosages are identical to the maximum tolerable dose (MTD) which was determined beforehand for each test substance in an extra experiment with nude mice.
The two tumor diameters are measured for each mouse twice a week, and the individual tumor area is calculated using the abovementioned formula.
Evaluation:
The relative tumor size VR and T/C ~ are calculated as above.
The statistical significance of the antitumor effect is determined using the Wilcoxon U test. The relative tumor sizes in the treated group are compared with the cor-3~:~
responding relative sizes in the control group on the same day of the experiment. The change in the tumor area is regarded as substance-speci~ic only if it is statisti-cally significant with p less than O . 05 .
Result:
Table 4 5~e Tm~r Dose Reyime Acti~ity (day 9);
(T/C %) .
Xylan poly- B 17 15 mg~bg QD9 x i.v. 61.4 ~- (~aan sulfate bron~
(n=9) camL%~) 20 mg/hg QD9 x i.v. 68.
Claims (17)
1. The use of a compound of the formula I
I
in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixtures with a mean molecular weight of about 1,000 to 20,000 Dalton, preferably of about 5,000 to 12,000 Dalton, especially about 6,000 Dalton (? = 9), or of one of the pharmacologically acceptable acid addition salts thereof, for the preparation of a pharmaceutical for the therapy of disorders based on uncontrolled and undifferentiated cell growth.
I
in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixtures with a mean molecular weight of about 1,000 to 20,000 Dalton, preferably of about 5,000 to 12,000 Dalton, especially about 6,000 Dalton (? = 9), or of one of the pharmacologically acceptable acid addition salts thereof, for the preparation of a pharmaceutical for the therapy of disorders based on uncontrolled and undifferentiated cell growth.
2. The use as claimed in claim 1 for the preparation of a pharmaceutical for the therapy of carcinomas or sarcomas.
3. The use as claimed in claim 1 for the preparation of a pharmaceutical for the therapy of renal carcinoma, mammary carcinoma, prostate carcinoma, pulmonary carcinoma, ovarian carcinoma, colon carcinoma, pancreatic carcinoma, rhabdomyosarcoma, melanoma or psoriasis.
4. The use of a compound as claimed in claim 1 for the preparation of a pharmaceutical acting to inhibit oncogene-encoded kinases and/or growth factor receptor tyrosine kinases.
5. A pharmaceutical composition for use in the treatment of disorders based on uncontrolled and undifferentiated cell growth comprising a compound of the formula I
I
in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixtures with a mean molecular weight of about 1,000 to 20,000 Dalton, preferably of about 5,000 to 12,000 Dalton, especially about 6,000 Dalton (? = 9), or of one of the pharmacologically acceptable acid addition salts thereof, and one or both of pharmaceutically acceptable auxiliaries and excipients.
I
in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixtures with a mean molecular weight of about 1,000 to 20,000 Dalton, preferably of about 5,000 to 12,000 Dalton, especially about 6,000 Dalton (? = 9), or of one of the pharmacologically acceptable acid addition salts thereof, and one or both of pharmaceutically acceptable auxiliaries and excipients.
6. A pharmaceutical composition as claimed in claim 5, for use in the treatment of carcinomas or sarcomas.
7. A pharmaceutical composition as claimed in claim 5, for use in the treatment of renal carcinoma, mammary carcinoma, prostate carcinoma, pulmonary carcinoma, ovarian carcinoma, colon carcinoma, pancreatic carcinoma, rhabdomyosarcoma, melanoma or psoriasis.
8. A pharmaceutical composition as claimed in claim 5, for use in inhibiting one or both of oncogene-encoded kinases and growth factor receptor tyrosine kinases.
9. The use of a compound of the formula I
I
in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixtures with a mean molecular weight of about 1,000 to 20,000 Dalton, preferably of about 5,000 to 12,000 Dalton, especially about 6,000 Dalton (? = 9), or of one of the pharmacologically acceptable acid addition salts thereof, in the treatment of disorders based on uncontrolled and undifferentiated cell growth.
I
in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixtures with a mean molecular weight of about 1,000 to 20,000 Dalton, preferably of about 5,000 to 12,000 Dalton, especially about 6,000 Dalton (? = 9), or of one of the pharmacologically acceptable acid addition salts thereof, in the treatment of disorders based on uncontrolled and undifferentiated cell growth.
10. The use of a compound as claimed in claim 9 in the treatment of carcinomas and sarcomas.
11. The use of a compound as claimed in claim 9 in the treatment of renal carcinoma, mammary carcinoma, prostate carcinoma, pulmonary carcinoma, ovarian carcinoma, colon carcinoma, pancreatic carcinoma, rhabdomyosarcoma, melanoma or psoriasis.
12. The use of a compound as claimed in claim 9 in inhibiting one or both of oncogene-encoded kinases and growth factor receptor tyrosine kinases.
13. A compound of the formula I
I
in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixtures with a mean molecular weight of about 1,000 to 20,000 Dalton, preferably of about 5,000 to 12,000 Dalton, especially about 6,000 Dalton (? = 9), or of one of the pharmacologically acceptable acid addition salts thereof, for use in the treatment of disorders based on uncontrolled and undifferentiated cell growth.
I
in which n is an integer from 1 to 20, preferably 8 to 10, especially 9, or of mixtures with a mean molecular weight of about 1,000 to 20,000 Dalton, preferably of about 5,000 to 12,000 Dalton, especially about 6,000 Dalton (? = 9), or of one of the pharmacologically acceptable acid addition salts thereof, for use in the treatment of disorders based on uncontrolled and undifferentiated cell growth.
14. A compound of the formula I as claimed in claim 13 for use in the treatment of carcinomas and sarcomas.
15. A compound of the formula I as claimed in claim 13 for use in the treatment of renal carcinoma, mammary carcinoma, prostate carcinoma, pulmonary carcinoma, ovarian carcinoma, colon carcinoma, pancreatic carcinoma, rhabdomyosarcoma, melanoma or psoriasis.
16. A compound of the formula I as claimed in claim 13, for use in inhibiting one or both of oncogene-encoded kinases and growth factor receptor tyrosine kinases.
17. The use of a compound as claimed in claim 1 and substantially as described herein.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE3917982A DE3917982A1 (en) | 1989-06-02 | 1989-06-02 | USE OF XYLANPOLYHYDROGENSULFATES FOR THE THERAPY OF CELL PROBLEMATION-RELATED DISEASES |
| DEP3917982.6 | 1989-06-02 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2018133A1 true CA2018133A1 (en) | 1990-12-02 |
Family
ID=6381904
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002018133A Abandoned CA2018133A1 (en) | 1989-06-02 | 1990-06-01 | Use of xylan polyhydrogensulfates for the therapy of disorders based on cell proliferation |
Country Status (7)
| Country | Link |
|---|---|
| EP (1) | EP0400586A1 (en) |
| JP (1) | JPH0320225A (en) |
| KR (1) | KR910000170A (en) |
| AU (1) | AU5617090A (en) |
| CA (1) | CA2018133A1 (en) |
| DE (1) | DE3917982A1 (en) |
| PT (1) | PT94211A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11274165B2 (en) | 2017-02-28 | 2022-03-15 | Oji Holdings Corporation | Pentosan polysulfate, pharmaceutical composition, and anticoagulant |
| US11278485B2 (en) | 2017-05-31 | 2022-03-22 | Oji Holdings Corporation | Moisturizing topical preparation |
| US11286272B2 (en) | 2016-08-31 | 2022-03-29 | Oji Holdings Corporation | Production method for acidic xylooligosaccharide, and acidic xylooligosaccharide |
| US11312790B2 (en) | 2016-08-31 | 2022-04-26 | Oji Holdings Corporation | Production method for pentosan polysulfate |
| US11344570B2 (en) | 2017-12-20 | 2022-05-31 | Oji Holdings Corporation | Pentosan polysulfate and medicine containing pentosan polysulfate |
| US11390693B2 (en) | 2017-09-12 | 2022-07-19 | Oji Holdings Corporation | Pentosan polysulfate and method for producing pentosan polysulfate |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0466315A3 (en) * | 1990-05-30 | 1992-07-01 | Larrian Gillespie | Compositions containing xylan sulphates for affecting growth factor function and for inhibiting fibroblast proliferation |
| NZ243082A (en) | 1991-06-28 | 1995-02-24 | Ici Plc | 4-anilino-quinazoline derivatives; pharmaceutical compositions, preparatory processes, and use thereof |
| GB9323290D0 (en) | 1992-12-10 | 1994-01-05 | Zeneca Ltd | Quinazoline derivatives |
| US6582919B2 (en) | 2001-06-11 | 2003-06-24 | Response Genetics, Inc. | Method of determining epidermal growth factor receptor and HER2-neu gene expression and correlation of levels thereof with survival rates |
| EP2220054A2 (en) | 2007-10-29 | 2010-08-25 | Natco Pharma Limited | Novel 4-(tetrazol-5-yl)-quinazoline derivatives as anti cancer agents |
| FI126211B (en) * | 2012-11-09 | 2016-08-15 | Montisera Ltd | Wood-based hemicelluloses for use in treating symptoms and diseases of the lower urinary tract |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2543145B1 (en) * | 1983-03-24 | 1986-10-17 | Sanofi Sa | NOVEL XYLAN SULPHATES, THEIR PREPARATION PROCESS AND THEIR ANTI-THROMBOTIC AND HYPOLIPEMIC ACTIVITY |
| DE3601136A1 (en) * | 1986-01-16 | 1987-07-23 | Max Planck Gesellschaft | INHIBITORS OF REVERSE TRANSCRIPTASE FOR PROPHYLAXIS AND THERAPY OF RETROVIRUS INFECTIONS IN MAMMALS |
-
1989
- 1989-06-02 DE DE3917982A patent/DE3917982A1/en not_active Withdrawn
-
1990
- 1990-05-30 EP EP19900110203 patent/EP0400586A1/en not_active Withdrawn
- 1990-05-31 PT PT94211A patent/PT94211A/en not_active Application Discontinuation
- 1990-05-31 AU AU56170/90A patent/AU5617090A/en not_active Abandoned
- 1990-05-31 KR KR1019900007944A patent/KR910000170A/en not_active Application Discontinuation
- 1990-06-01 CA CA002018133A patent/CA2018133A1/en not_active Abandoned
- 1990-06-01 JP JP2141746A patent/JPH0320225A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11286272B2 (en) | 2016-08-31 | 2022-03-29 | Oji Holdings Corporation | Production method for acidic xylooligosaccharide, and acidic xylooligosaccharide |
| US11312790B2 (en) | 2016-08-31 | 2022-04-26 | Oji Holdings Corporation | Production method for pentosan polysulfate |
| US11274165B2 (en) | 2017-02-28 | 2022-03-15 | Oji Holdings Corporation | Pentosan polysulfate, pharmaceutical composition, and anticoagulant |
| US11278485B2 (en) | 2017-05-31 | 2022-03-22 | Oji Holdings Corporation | Moisturizing topical preparation |
| US11390693B2 (en) | 2017-09-12 | 2022-07-19 | Oji Holdings Corporation | Pentosan polysulfate and method for producing pentosan polysulfate |
| US11344570B2 (en) | 2017-12-20 | 2022-05-31 | Oji Holdings Corporation | Pentosan polysulfate and medicine containing pentosan polysulfate |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0320225A (en) | 1991-01-29 |
| DE3917982A1 (en) | 1990-12-06 |
| EP0400586A1 (en) | 1990-12-05 |
| AU5617090A (en) | 1990-12-06 |
| KR910000170A (en) | 1991-01-29 |
| PT94211A (en) | 1991-02-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2018133A1 (en) | Use of xylan polyhydrogensulfates for the therapy of disorders based on cell proliferation | |
| Piantelli et al. | Tamoxifen and quercetin interact with type II estrogen binding sites and inhibit the growth of human melanoma cells | |
| Presta et al. | Purine analogue 6-methylmercaptopurine riboside inhibits early and late phases of the angiogenesis process | |
| Iyer et al. | Targeting TGF beta signaling for cancer therapy | |
| JP2005532805A5 (en) | ||
| Peterson et al. | Interleukin-11 induces intestinal epithelial cell growth arrest through effects on retinoblastoma protein phosphorylation | |
| Garona et al. | Preclinical efficacy of [V4Q5] dDAVP, a second generation vasopressin analog, on metastatic spread and tumor-associated angiogenesis in colorectal cancer | |
| SK13912002A3 (en) | Combination therapies with vascular damaging activity | |
| Favoni et al. | Preclinical studies identify novel targeted pharmacological strategies for treatment of human malignant pleural mesothelioma | |
| Amorino et al. | Radiopotentiation by the oral platinum agent, JM216: role of repair inhibition | |
| CN114761006A (en) | Methods of treating cancer resistant to kinase inhibitors | |
| Berns et al. | Antiproliferative effects of suramin on androgen responsive tumour cells | |
| NZ537759A (en) | Combination of chemotherapeutic drugs for increasing antitumor activity | |
| Rusnati et al. | Thrombospondin‐1/HIV‐1 Tat protein interaction: modulation of the biological activity of extracellular Tat | |
| US20030045550A1 (en) | Bicyclic aromatic chemokine receptor ligands | |
| US20110263525A1 (en) | Drug composition cytotoxic for pancreatic cancer cells | |
| DE69305348T2 (en) | Protein tyrosine phosphatase alpha inhibitors for tumor treatment | |
| Badalamenti et al. | Soft tissue sarcomas in the precision medicine era: new advances in clinical practice and future perspectives | |
| Detchou et al. | Interleukin 6 and cancer resistance in glioblastoma multiforme | |
| EP1651227B1 (en) | Pharmaceutical compositions comprising azd 2171 and zd 6126 and uses thereof | |
| US7297779B2 (en) | Colon cancer metastasis inhibitor | |
| KR101385488B1 (en) | Inhibition of osteolytic lesions by SRC kinase inhibitors | |
| Honma et al. | Induction by some protein kinase inhibitors of differentiation of a mouse megakaryoblastic cell line established by coinfection with Abelson murine leukemia virus and recombinant SV40 retrovirus | |
| Retz et al. | New HIV-drug inhibits in vitro bladder cancer migration and invasion | |
| Iwamoto et al. | Use of a reconstituted basement membrane to study the invasiveness of tumor cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Dead |