Agent eliciting antitumor effect

FIELD: chemistry of peptides, medicine, oncology, pharmaceutical chemistry.

SUBSTANCE: invention relates to the development of medicinal agent of peptide nature eliciting an antitumor effect and can be used in treatment of endocrine and hormone-dependent tumors. Agent represents peptide of the general formula: . Invention provides enhancement of the therapeutic effect and reducing toxicity.

EFFECT: valuable medicinal properties of agent.

3 cl, 4 tbl, 2 ex

 

The invention relates to pharmaceutical chemistry, in particular to the creation of medicines peptide having anti-tumor activity, and can be used to treat tumors, including endocrine and hormone-dependent.

In the modern practice of anticancer chemotherapy uses a very wide range of drugs, such as alkylating agents, antimetabolites, antitumor antibiotics, hormones and their antagonists, enzymes. Widely used cytostatics sarcolysin, cyclophosphamide, malasan, derived nitrosodiethylamine, antimetabolites methotrexate, mercaptopurine, fluorouracil, Ftorafur, among antitumor antibiotics, dactinomycin, rubomycin, doxorubicin, karminomitsin and others.

For the treatment of hormone-dependent tumors often use hormones, particularly androgens, estrogens, corticosteroids, and hormone antagonists. So, for the treatment of prostate cancer used estrogen, as in this case shows suppression actions in the body of androgens or increased activity of estrogen. Androgens are used, when shown an increase in the activity of androgens or reducing the activity of estrogen, such as breast cancer.

For the treatment of malignant lymphoma and acute is Anosov use glucocorticoids.

In modern medical Oncology practice for the treatment of hormone-dependent tumors using drugs such as anastrozole, megestrol, medroxyprogesterone, tamoxifen, flutamide and other (RLS Russia, encyclopedia of medicine, volume 9, 2002). But these medicines, in addition to serious side effects, and have absolute contraindications, such as liver and kidney failure, thrombosis, diseases of the thyroid gland and heart, child, pregnancy.

In recent decades is an active search for anticancer drugs peptide having a high specific activity and at the same time with no serious contraindications.

Renowned pharmaceutical composition having antitumor activity, containing β-geptilglycoside-N-acetylmuramyl-L-alanyl-D-isoglutamine in liposomal form for oral and intravenous (patent RU 2121363, 1997), the composition has a low toxicity.

Known antitumor agent representing complex drug hematoporphyrin derivative and tumor necrosis factor alpha in a molar ratio (Patent RU 2136278, 1998), the tool has a low toxicity.

Known Pentapeptide with antitumor activity is translated in the form of hydrochloride M 2Val-Val-MVal-Pro-Pro-NH-Bzl Hcl, where M is the metal, Bzl is benzyl (patent RU 2153504, 1995).

Known antitumor agent, representing nontoxic Hexapeptide of the formula Leu-Val-Val-Tyr-Pro-Trp (patent RU 2067870, 1993).

Known pharmaceutical composition based on peptides, possessing antiproliferative activity against tumors (patent RU 2156774, 1995).

More and more wide application in clinical therapy of tumors, especially of endocrine tumors, finds sandostatin, a drug on the basis of cyclic oktapeptid manufactured by Sandoz (Nfrl and others, as well. "Modern Oncology", vol. 2, No. 1, 2000).

The pharmacological spectrum of action and specific antitumor activity sandostatin is closest to the claimed means, therefore, selected by the applicant as a prototype.

The present invention is the creation of a domestic anti-cancer drugs with a high therapeutic effect, a wide spectrum of antitumor activity and low toxicity.

The problem is solved in that the proposed agent with antitumor activity representing a peptide of General formula:

where

R - tert-butyloxycarbonyl, R1- tetrahydropyranyl, R2-Nε- benzyloxycarbonyl, R3- me is Il (I);

R-N, R1- tetrahydropyranyl, R2-Nε- benzyloxycarbonyl, R3- methyl (II);

R-N, R1-H, R2-N, R3- methyl (III);

R - tert-butyloxycarbonyl, R1- tetrahydropyranyl, R2-Nε- benzyloxycarbonyl, R3-S - tetrahydropyrimidine methyl ether (IV).

The technical result to be obtained by the use of the invention is expressed in expanding Arsenal of anticancer agents, reduce toxicity, improve therapeutic effect.

The synthesis of the claimed means were carried out by classical methods of peptide chemistry. To lock the side chain of lysine used carbobenzoxy (Z) group of cysteine - tetrahydropyranyl (Thp), for a temporary protection Nαfunctions tert-butyloxycarbonyl (BOC) group.

To increase the resistance to tissue enzymes in the composition of peptides introduced non-natural amino acid is D-tryptophan.

The resulting peptides were characterized by optical rotation angle and the data of elemental analysis, thin-layer chromatography, high performance liquid chromatography, data, NMR spectroscopy and amino acid analysis.

The invention is illustrated by the following examples

Example 1

In the example of the synthesis of the hydrochloride of the methyl ester of S-tetrahydropyrimidine the Anil-D-tryptophyl-N ε-carbonisation

HCl·H-Cys(Thp)-Phe-D-Trp-Lys(Z)-Thr-OMe (II)

Pentapeptide (II) was obtained by removal of the BOC-protective group from Pentapeptide (I) 3,4 N Hcl in ethyl acetate at room temperature for 20 minutes. In these conditions has not been observed adverse reaction removal of the Z-group with Nεfunction-lysine. Output (II) was 98%.

Example 2

The peptide obtained by the described method was tested on adequate models: adenocarcinoma of the prostate of the Dunning rat Copenhagen and breast cancer RM-1 in Nude mice. In the experiment with rats the drug was administered in the dose of 100 g/kg for 30 days in mice in a dose of 100 g/kg for 21 days.

As shown by experimental data, the peptide exhibits equal antitumor effect on adenocarcinoma of the prostate of the Dunning R3327-H rats and cancer of the breast RM-1 Nude mice. The peptide inhibits the growth of these tumors at 63-65%.

Experimentally it was found that the claimed product has a broad spectrum of antitumor activity, and it is now on transplantable tumors: breast adenocarcinoma CA-755, melanoma b-16, lung adenocarcinoma Lewis LLC, adenocarcinoma of the prostate of the Dunning R-3327-H, breast cancer RM-1 Nude mice, DMBA - induced mammary tumors.

Were Prov is found tests of funds.

1. The study of toxicity.

2. The study of specific antitumor activity.

1. The toxicity study of the proposed drug compounds I, II, III, IV were performed on mice-hybrids BDF1and outbred rats females. All connections do not show toxic effects on mice at single doses of 60-80 mg/kg (total dose : 300-400 mg/kg). In terms of human daily dose of some 300,000-400,000 g, which 50-65 times used with inoperable cancer of the pancreas high doses of sandostatin (6000 mg/day). In outbred rats all connections also do not show toxic effects in exchange dose of 6000 mg/kg

The data presented show that the proposed remedy does not show toxicity when used even in high doses.

2. The study of antitumor activity was carried out on adequate models. And effectiveness of treatment was the inhibition of tumor growth (SRW). Antitumor activity was studied on adenocarcinoma of the prostate of 3327 Dunning H rat Copenhagen. The data presented in table 1.

As can be seen from table 1, compounds I, III, IV for adenocarcinoma of the prostate of the Dunning R3327-H rats antineoplastic action: compound I inhibits tumor growth by 55%, the compound III - 66%, compound IV - 46%.

Antitumor activity of the proposed drug was studied on DMBA-induced mammary tumors in rats in comparison with sandostatin. The test drugs were administered subcutaneously at a dose of 100 g/kg for 30 days. The data presented in table 2.

As can be seen from table 2, all compounds exhibit high antitumor effect. therapeutic effect of compound I is 91% SRAW. The compound III maximum therapeutic effect is equal to 75% of the solid from compound IV - 79% SRAW, and sandostatin - 68% SRAW.

Antitumor activity of the proposed drug also studied in Nude mice with subcutaneously inoculated breast cancer RM-1 in comparison with sandostatin. The compound was administered subcutaneously at a dose of 100 g/kg for 21 days. The data presented in table 3. As can be seen from table 3, the infusion of connections maximum therapeutic effect amounted to compound I - 47% SRAW, for compound II - 63% SRAW and exceeded the antitumor effect of sandostatin (39% SRW). Directly after the treatment, therapeutic effect was maintained and accounted for compound I - 45% SRAW, for compound II - 55% SRAW, for sandostatin - 40% SRAW.

In the experiment shown a statistically significant anti-tumor effect of the proposed drug on transplantable gormononezawisimy tumors mammary adenocarcinoma CA-755, lung adenocarcinoma Lewis LLC and melanoma b-16

Study drugs were administered at doses of 60-80 mg/kg subcutaneously for 5 days.

Dan is s presented in table 4. As can be seen from table 4, the maximum TRO on mammary adenocarcinoma CA-755 91%, lung adenocarcinoma Lewis - 97% and melanoma b-16 - 85%.

Thus, the claimed product has a high antitumor effect on a wide range of tumors, including hormone-dependent, and low toxicity, which will enable it to take its rightful place in oncological practice.

Table 1

Antitumor activity of compounds I, III, IV adenocarcinoma of the prostate of 3327 Dunning H rat Copenhagen
MedicationDose (m kg/kg) *Maximum SRW, % **
I10055
III10066
IV10046
Notes: * connection was administered daily for 30 days,

** p<0,05

91
Table 2

Antitumor activity of compounds I, III, IV, DMBA-induced mammary tumors in rats in comparison with sandostatin
MedicationDose (g/kg)*Maximum SRW, % **
I100
III10075
IV10079
Sandostatin10068
Notes: * connection was administered daily for 30 days,

** p<0,05

Induction of tumors was carried out four times weekly intravenous administration of DMBA (dimethylbenzanthracene) in water-fat emulsion in a total dose of 8 mg per rat.

Table 4

Antitumor activity of compounds I, III, IV on transplantable tumors of mice
ModelDose (mg/kg) *Maximum SRW, % **
CA-7558091
LLC7097
B-166085
Notes: * connection was administered daily for 5 days,

** p<0,05

LITERATURE

1. Romy Lea Topper, Pharm D.Ocreotide: Drug review and clinical indications. The cancer bulletin, 1989, 41, 5, 334-335.

2. Garin A. M. the Principles and possibilities of modern endocrine therapy of tumors. M, 2000, 82-85.

3. Agthony L.B., Krozely M.G., Jonhson D.H., Shaff M.I., Hande K.R., Oates J.A.Somatuline is antitumor efficacy and phase I trial in neuroendocrine tumors. Proc. Annu MeetAm Soc Clin Oncol, 1992, 11:A.

4. Jean-Christophe Souget. Long-acting somatostatin analogues in pancreatic islet cell carcinoma. Horm. Res., 1989, 32, 74-76.

5. Weckbecker G., Raulf f, Stolz C., Bruns C. Somatostatin analogs for diagnosis and treatment of cancer. Pharm. Ther. Vol. 60, p. 245-264, 1993.

6. Cavanac et al. Somatostatin analogue composition and in treating beastm cancer U.S.P.

7. Nfrl, Tnguatky. Possibilities of application of sandostatin in cancer patients. Modern Oncology, 2, 1, 2000.

1. The agent with antitumor activity that contains a peptide and a physiologically acceptable solvent, characterized in that it contains peptide is a linear peptide of General formula:

where

R - tert-butyloxycarbonyl or N,

R1- tetrahydropyranyl or N,

R2-Nε- benzyloxycarbonyl or N,

R3- O-methyl or S-tetrahydropyrimidine methyl ester.

2. The tool according to claim 1, characterized in that it is intended for parenteral administration contains the active ingredient in a dose of 0.1-100.0 mg/kg

3. The tool according to claim 1, characterized in that it is intended for parenteral administration, made in liposomal form and contains the active substance at a dose of 0.05-100 mg/kg of body weight.



 

Same patents:

FIELD: biochemistry, medicine, pharmacy.

SUBSTANCE: leaves of Mikania micrantha, Mikania scandens and Mikania cordata are milled and dries and a solvent - toluene, ethyl acetate, their mixture, mixture of toluene with acetone, mixture of ethyl acetate with heptane or mixture of heptane with acetone is added in the ratio (7:3)-(3:7), mixture is filtered and concentrated up to 2.5-10% as measure for dry extract. Then extract is contacted with a mixture containing 10-50% of methanol or ethanol in water, an aqueous-alcoholic phase is washed out with n-hexane or heptane and alcohol is removed. An aqueous phase is extracted with ethyl acetate, prepared phase is dried, solvents are evaporated and dry extracted is purified. Dry extract prepared from leaves is dissolved in ethyl acetate and hydrogenated at 10-35°C in the presence of hydrogenation catalyst under pressure 1-2 atm. After hydrogenation dihydromikanolid is crystallized. Mikanolid and dihydromikanolid are used for treatment proliferative diseases and parasitic diseases. Also, invention relates to a medicinal agent containing above indicated vegetable extract. Invention provides realization of indicated designation.

EFFECT: valuable properties of agents.

12 cl, 3 tbl, 3 ex

FIELD: organic chemistry, medicine, oncology, pharmacy.

SUBSTANCE: invention relates to pharmaceutical composition comprising the known antibiotic eliciting an antitumor activity and derivative of hydroxamic acid of the formula (1) wherein the weight ratio of two active agents is (1:50)-(50:1), respectively. The composition reduces adverse effects and elicits the enhanced antitumor effect.

EFFECT: improved valuable medicinal properties of composition.

4 cl, 1 tbl, 5 ex

FIELD: organic chemistry, biochemistry.

SUBSTANCE: invention relates to epothilones with modified thiazole substituent, methods for production thereof and pharmaceutical composition capable of cell growth inhibiting containing the same. Claimed compounds have general formula I , wherein P-Q represents double carbon bond or epoxy; R represents H, C1-C6-alkyl; G represents ; R1 represents and ; G1 and G2 represent hydrogen; G3 represents O, S, and NZ1; G4 represents H, optionally substituted C1-C6-alkyl, OZ2, Z2C=O and Z4SO2; G5 represents halogen, N3, CN, NC, heteroaryl containing nitrogen or oxygen, and heterocycle; G6 represents H, C1-C6-alkyl, or OZ5, wherein Z5 represents H, C1-C6-alkyl; G9 represents oxygen; Z1 represents H, optionally substituted C1-C6-alkyl, optionally substituted acyl; Z2 represents optionally substituted C1-C6-alkyl or aryl; Z4 represents optionally substituted aryl.

EFFECT: new epothilones capable of cell growth inhibiting.

19 cl, 39 ex

FIELD: medicine, oncology.

SUBSTANCE: the present innovation should be applied at adjuvant chemotherapy in case of tumors of central nervous system. Moreover, while carrying out lumbar puncture it is necessary to perform catheterization of subarachnoidal space. Moreover, one should daily sample liquor at the quantity of 10 ml to be incubated with chemopreparation in vitro for 30 min at 38 C. One should daily introduce chemopreparations upon autoliquor through catheter during the whole period of therapy course. The method enables to choose any mode and duration of endolumbar chemotherapy at its decreased toxicity.

EFFECT: higher efficiency of chemotherapy.

1 ex

Antitumor agent // 2253446

FIELD: drugs, medicine.

SUBSTANCE: invention relates to application of 2-METHYL-4-chlorophenoxyacetic acid tris-(2-hydroxyethyl)ammonia salt, which represents known microorganism living function stimulator, as antitumor agent and metastasis inhibitor. Present invention makes it possible to produce drugs for treatment cancers, in particular hepatoma, melanocytoma, lymphadenoma, etc.

EFFECT: new drug for cancer treatment.

2 tbl, 2 ex

FIELD: pharmaceutical industry, in particular new bioactive chalcones.

SUBSTANCE: invention relates to new chalcones of formula I

, pharmaceutically acceptable salts or solvates thereof, wherein Ar is optionally substituted C5-C10-carbocycle group or 5- or 6-membered heterocycle group having sulfur atom in cycle, and Ar substituents are selected independently from Cl, Br, F, CN, SCH3 and OR10, wherein R10 is linear or branched C1-C6-hydrocarbon; R is OH or R10; R2 and R3 are independently phenyl, saturated linear or branched C1-C6-hydrocarbon, or R2 and R3 together with carbon atom attached thereto form 5- or 6-membered carbocycle group with the proviso, that in compounds where R is OH and both R2 and R3 are methyl, Ar is not phenyl, 4-chlorophenyl, 4-chlorophenyl, 4-methylphenyl, 2-chlorophenyl, 3,4-dimethoxyphenyl, or 4-methoxyphenyl. Also disclosed are drug component for treatment or prophylaxis of neoplasm and pharmaceutical compositions with antiproliferation effect based on compounds of formula I.

EFFECT: new chalcone derivatives with value bioactive action.

26 cl, 2 tbl, 22 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: the suggested composition includes compounds of natural ethers in cancer of mammalian soft tissues (including, the man), for example, in case of diseases of mammary gland. Both dosage and frequency of introduction depending upon a certain symptomatics could be decreased up to the level at which it is necessary to maintain improved level. At weakening the symptoms up to desired level one should stop therapy immediately. At any relapse of disease symptoms patients could need periodic therapy upon lasting basis.

EFFECT: higher efficiency of application.

2 cl, 3 ex, 3 tbl

FIELD: medicine.

SUBSTANCE: the present innovation includes polychemotherapy and radiation therapy. Moreover, polychemotherapy should be carried out by the following scheme: on the 1st and the 8th d of the first and the third courses it is necessary to introduce doxorubicin, cyclophosphan, vincristine, and since the 1st to the 14th d - procarbazine and prednisolone; moreover, on the 1st and the 8th d of the second and the fourth courses one should introduce doxorubicin, bleomycin, vinblastine, dacarbazine. The method enables to decrease the quantity of late therapeutic complications, improves the results of relapse-free, total tumor-specific survival rate and decreases the number of polychemotherapeutic cycles.

EFFECT: higher efficiency of therapy.

2 ex

FIELD: medicine, oncology.

SUBSTANCE: the present innovation deals with treating oncological diseases. It is suggested to apply bisdioxopiperazine (previously known as cardioprotector) to either treat or prevent tissue lesions caused due to sporadic transudation of cytotoxic poison for topoisomerase II (represented by anthracyclines, etoposide, teniposide, mitoxantrone daunorubicin, doxorubicin, etc.), medicinal remedies and pharmaceutical set of the same indication. It is, also, suggested to apply the method to treat or prevent tissue lesions caused by sporadic transudation of topoisomerase II poison. BisdioxopiperazineICRF-187 has impact due to catalytic inhibiting topo II. Signs for possible transudation of topoisomerase II poison (of local toxicity) usually include the availability of acute pain, erythema, development of ulcerations in area of transudation; due to the action of ICRF-187 the quantity of wounds is reduced, or the development of side effects is not observed.

EFFECT: higher efficiency of therapy.

59 cl, 12 dwg, 13 ex, 10 tbl

FIELD: genetic engineering, in particular genes for cell cycle controlling point.

SUBSTANCE: polynucleotide encoding rad3 polypeptide ATR homologue is cloned into expression vector, having functionality in eucariotic cells. Polypeptide of rad3 polypeptide ATR homologue is obtained by cultivation of eucariotic cell culture, transformed by vector. Monoclonal antibody to rad3 polypeptide ATR homologue is obtained by hybridoma technologies. Polyclonal antibodies are obtained by inoculation of rad3 polypeptide ATR homologue in host animal. Polynucleotide presence in animal tissue sample is detected by contacting of this sample containing DNA or RNA with polynucleotide encoding rad3 polypeptide ATR homologue under hybridization conditions. Polypeptide in biological sample is detected by sample contact with monoclonal or polyclonal antibodies. Substances having anticancer activity are screened on the base of reduced activity of ATR polypeptide on substrate or reduced chelating of ATR homologue in presence of candidate substance. Present invention makes it possible to produce human or S.pombe rad3 polypeptide ATR homologue and is useful in investigation ATR role as gene for cell cycle controlling point in cell culture in vivo or in vitro.

EFFECT: new anticancer substances.

24 cl, 1 dwg

FIELD: medicine.

SUBSTANCE: peptide of the following formula: X-Pro-Gly-P, where X = Thr-Lys-Pro-Arg-; Lys-pro-Arg-; pro-Arg-; Arg-, being of untiulcerous activity. They should be applied at intraperitoneal injection at the dosage of 0.58-3.20 mcM g/kg for preventing and treating ulcers of gastro-intestinal tract.

EFFECT: higher efficiency and prophylaxis.

4 dwg, 5 ex

FIELD: genetic engineering, medicine.

SUBSTANCE: invention relates to T-cell receptor sequence being detected in patients with extended sclerosis and is useful in diagnosis and therapy. Oligonucleotide including sequence which represents or is derived from 5'-CTAGGGCGGGCGGGACTCACCTAC-3' or nucleotide sequence being fully complementary thereto. Oligonucleotide together with nuclear acid including nearly 15-30 oligonucleotides, which doesn't comprise oligonucleotide sequence and presents in region from Vβ to Jβ of Vβ13.1 gene in T-cell Vβ13.1-subgroup, wherein oligonucleotide and nuclear acid sequences don't present in the same chain of pair sequences of Vβ13.1 gene, is used in Vβ13.1 gene part amplification. In method for detection of LGRAGLTY motive, which is present in T-cell receptors of T-cell Vβ13.1-subgroup, oligonucleotide is used in combination with labeling particle. Once LGRAGLTY motive is detected, development monitoring and treatment are carried out by removing of LGRAGLTY motive-containing peptide.

EFFECT: simplified methods for detection of LGRAGLTY motive in T-cell receptors and treatment of patients with extended sclerosis.

21 cl, 7 dwg, 3 tbl, 3 ex

FIELD: pharmaceutical chemistry, medicine.

SUBSTANCE: in the suggested composition one should apply heptapeptide of Met-Glu-His-Phe-Pro-Gly-Pro sequence (heptapeptide A) for treating ischemic insult due to introducing 2 drops of compositions into each nasal canal 5-6 times daily for 10 d at disease of average severity degree, and in case of severe degree - per 3 drops of the present composition into each nasal canal 7 times daily for 10 d. The present innovation provides increased efficiency at decreased concentration of heptapeptide without any side effects.

EFFECT: higher efficiency of therapy.

2 cl, 6 dwg, 8 ex, 5 tbl

FIELD: medicine, phthisiology, anesthesiology.

SUBSTANCE: during the day of operation one should perform autohemotransfusion, then introduce epocrine intravenously by drops at the dosage of 50-200 U/kg patient's body weight; next day after interference one should inject epocrine subcutaneously at the dosage of 25-100 U/kg; at hematocrit level being below 35% 48 h after operation it is necessary to repeat subcutaneous injection of the above-mentioned preparation at the dosage not exceeding 50 U/kg. The present innovation favors hemopoiesis stimulation in postoperational period, that, in its turn, accelerates postoperational rehabilitation in patients of this group and enables, also, to avoid allotransfusions being dangerous because of immunoconflicting reactions.

EFFECT: higher efficiency of compensation.

1 ex, 1 tbl

FIELD: medicine, cardiology.

SUBSTANCE: the suggested method should be performed at the background of medicinal therapy with preparations out of statins group, tevetene, polyoxidonium and conducting seances of plasmapheresis by removing 800 ml plasma twice weekly with N 5 due to additional intramuscular injection of immunophan 0.005%-1.0 with N 10 and fluimucyl 300 mg intravenously daily with N 5-10, total course of therapy lasts for 2 mo. The method provides modulation of leukocytic functional activity, moreover, due to altered cytokine profile and, thus, through disintegration of protein-lipid complexes participating in the development of atherosclerotic platelets.

EFFECT: higher efficiency of therapy.

3 ex

The invention relates to pharmaceutical industry and relates to the preparation of dalargin for injection, used as a medicine in the acute gastric ulcer and duodenal ulcer, acute pancreatitis and pancreonecrosis

The invention relates to medicine and relates to a medicinal product and method of treatment of neurological diseases in children from infant age up to 7 years

The invention relates to the field of biotechnology, specifically to peptides active in the attachment, respectiveiy and detaching cells, and can be used to study activity in the attachment of cells, mediated by different proteins of the extracellular matrix, to develop peptides, adhesion, and also for medicinal purposes

FIELD: medicine, cardiology.

SUBSTANCE: the suggested method should be performed at the background of medicinal therapy with preparations out of statins group, tevetene, polyoxidonium and conducting seances of plasmapheresis by removing 800 ml plasma twice weekly with N 5 due to additional intramuscular injection of immunophan 0.005%-1.0 with N 10 and fluimucyl 300 mg intravenously daily with N 5-10, total course of therapy lasts for 2 mo. The method provides modulation of leukocytic functional activity, moreover, due to altered cytokine profile and, thus, through disintegration of protein-lipid complexes participating in the development of atherosclerotic platelets.

EFFECT: higher efficiency of therapy.

3 ex

Up!