KR20130018548A - Novel prodrugs of 5-(2,4-dihydroxy-5-isopropylphenyl)-n-ethyl-4-(5-methyl-1,2,4-oxadiazol-3-yl)isoxazole-3-carboxamide - Google Patents
Novel prodrugs of 5-(2,4-dihydroxy-5-isopropylphenyl)-n-ethyl-4-(5-methyl-1,2,4-oxadiazol-3-yl)isoxazole-3-carboxamide Download PDFInfo
- Publication number
- KR20130018548A KR20130018548A KR1020120082521A KR20120082521A KR20130018548A KR 20130018548 A KR20130018548 A KR 20130018548A KR 1020120082521 A KR1020120082521 A KR 1020120082521A KR 20120082521 A KR20120082521 A KR 20120082521A KR 20130018548 A KR20130018548 A KR 20130018548A
- Authority
- KR
- South Korea
- Prior art keywords
- formula
- compound
- methyl
- oxadiazol
- ethylcarbamoyl
- Prior art date
Links
- 0 CC(C)c(c(O*)c1)cc(-c([o]nc2*)c2-c2n[o]c(C)n2)c1O* Chemical compound CC(C)c(c(O*)c1)cc(-c([o]nc2*)c2-c2n[o]c(C)n2)c1O* 0.000 description 2
- FADKJNSSIWTVAI-SSDOTTSWSA-N CC([C@@H](CCCCN)N)=O Chemical compound CC([C@@H](CCCCN)N)=O FADKJNSSIWTVAI-SSDOTTSWSA-N 0.000 description 1
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
- A61K31/66—Phosphorus compounds
- A61K31/661—Phosphorus acids or esters thereof not having P—C bonds, e.g. fosfosal, dichlorvos, malathion or mevinphos
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
- C07F9/02—Phosphorus compounds
- C07F9/06—Phosphorus compounds without P—C bonds
- C07F9/08—Esters of oxyacids of phosphorus
- C07F9/09—Esters of phosphoric acids
Abstract
The present invention relates to novel compounds of formula (I). The compounds of formula (I) according to the invention are used as prodrugs of compounds of formula (II) which are parent compounds, which have valuable pharmacological properties.
The compound of formula (I) of the present invention has a solubility of 1,000 to 20,000 times or more compared to the compound of formula (II), which is a parent compound, and thus can increase bioavailability, and AUC 0 -120 min is 3 to 5 times and C max is greater than that of formula (II). It is absorbed in vivo at a concentration of 10 to 20 times, which greatly improves pharmacokinetic properties and can be used to treat various tumors, including ovarian cancer and gastric cancer.
Description
The present invention relates to compounds of formula (I)
<Formula I>
Where
R 1 and R 2 are both PO (OH) 2 or
The compound of formula I is 5- (2,4-dihydroxy-5-isopropylphenyl) -ene-ethyl-4- () which is a compound of formula II which inhibits HSP90 activity disclosed in WO 2011/102660. It has been found that it can be used as a prodrug of 5-methyl-1,2,4-oxadiazol-3-yl) isoxazole-3-carboxamide.
<Formula II>
Prodrugs are biologically inactive derivatives of the parent drug molecule that in most cases require spontaneous or enzymatic modification in the body to release the active drug and have improved delivery properties over the parent drug molecule. It has been shown that in molecules with physicochemical properties and optimal structural coordination to elicit the desired therapeutic response at the target site, it is not necessary to have the best molecular morphology and properties for delivery to its ultimate point of action. Typically, only a minimal amount of the dose administered reaches the target site, and because most agents also interact with sites other than the target, inefficient delivery can result in unwanted side effects. This fact that the effect characteristics of delivery and in-situ differ for many drug molecules is that bio-reversible chemical induction of the drug, ie prodrug formation, often results in substantial improvement in the overall efficacy of the drug. It's the basic reason for achieving this. Prodrugs are designed to solve problems based on pharmacokinetics associated with parent drug molecules.
Recently, several types of bio-reversible derivatives have been developed for use in designing prodrugs. It is most common to use esters containing carboxyl or hydroxyl functional groups as types for drugs. Also, prodrug derivatives such as peptides and 4-imidazolidinones ( Drugs) of the Future , 1991, 16 (5), 443-458), prodrug derivatives of N-oxides (US Pat. No. 5,691,336) are known.
Molecular chaperone, on the other hand, is a generic term for proteins that temporarily form complexes with target proteins to promote the formation of functional higher order structures of proteins. Molecular chaperones help protein folding and association, and have the activity of inhibiting aggregation.
In general, exposure of cells to a variety of environmental disorders, including heat, alcohol, heavy metals and oxidative disorders, typically results in cell accumulation in many chaperones known as heat shock proteins (HSPs). These heat shock proteins (HSPs) are classified into HSP70, HSP90, and HSP27 according to the molecular weight. They protect cells against the initial impairment of impairment and enhance the maintenance and recovery induction of impaired resistance. In addition, any heat shock protein (HSPs) regulate folding, degradation, integration and function of important cellular proteins.
Some diseases of humans can be caused by the consequences of misfolded proteins. For example, proteins that are misfolded in Alzheimer's, prion and Huntington's disease can cause protein aggregation, leading to neurodegenerative diseases (Tytell M. and Hooper PL, Emerging Ther. Targets (2001) 3796). In addition, heat shock proteins (HSPs), especially HSP90, affect tumors through activation of various client proteins involved in cell proliferation or apoptosis. It has been reported that treatment of diseases through inhibition of chaperone function in such pathological symptoms is possible, and in particular inhibition of the function of HSP90 is useful for tumor treatment (Moloney A. and Workman P., Expert Opin. Biol. Ther. (2002), 2 (1), 3-24]; [Choisis et al, Drug Discovery Today (2004), 9, 881-888).
HSP90 (Heat Shock Protein 90) consists of HSP90α, HSP90β, GRP94 and HSP75 / TRAP1, which accounts for about 1-2% of total cell protein mass. It is in the form of a dimer in the cell and is associated with various proteins called co-chaperones. HSP90 restores original folding or protein aggregation through interaction with many proteins whose original folding has been modified by external stresses such as thermal shock (Smith DF et al., Pharmacological Rev. 1998), 50, 493-513). HSP90 also plays an important role as a buffer against mutations through modification of inaccurate protein folding caused by mutations (Rutherford and Lindquist, 1998). Under physiological conditions, HSP90 binds to a variety of client proteins (e. G., EGFR R / HER2, Src, Akt, Raf, MEK, Bcr- Abl, Flt-3, p53 in mutant form, Akt, And plays a role in stabilizing the stability of the sequences of the survivin, CDK4, Plk, Wee1, VEGF-R, FAK, HIF-1, hTert and c-Met etc. . These client proteins have the ability to proliferate in the absence of growth factors (EGFR-R / HER2, Src, Akt, Raf, MEK, Bcr-Abl, Flt-3, etc.); ii) the ability to evade apoptosis (mutant forms of p53, Akt, survivin, etc.); iii) insensitivity to the proliferation stop signal (Cdk4, Plk, Wee1, etc.); iv) the ability to activate angiogenesis (VEGF-R, FAK, HIF-1, Akt, etc.); v) the ability to propagate without any replication limitations (hTert, etc.); vi) involvement in the ability to evade new metastases (c-Met) (Hanahan D. and Weinberg RA, Cell (2002), 100, 57-70). Thus, inhibition of HSP90 activity can inhibit tumor formation by these client proteins.
Currently known HSP90 inhibitors are the first reported compounds of the ansamycin group, in particular geldanamycin and herbimycin A. X-ray studies have shown that geldanamycin binds to the ATP site of the N-terminal domain of HSP90, where geldanamycin inhibits chaperone ATPase activity (Prodromou C. et al, Cell (1997) 90, 65-75). Currently, NIH and KOSAN Biosciences are under clinical trials for 17AAG derived from geldanamycin.
Radicicol is a natural HSP90 inhibitor (Roe SM et al., J. Med Chem. (1999), 42, 260-66). Radicocor is the best in vitro HSP90 inhibitor to date, but it is difficult to use in vivo because it is metabolically labile to sulfur-containing nucleophiles. The natural HSP90 inhibitor novobiocin binds to other ATP sites in the C-terminal domain of the protein (Itoh H. et al, Biochem J. (1999), 343, 697-703). PU3 ([Chiosis et al, Chem. Biol. (2001), 8, 289-299), a purine-based compound, was reported as an HSP90 inhibitor as a low molecular HSP90 inhibitor.
(WO 2004/072080), phthalazole derivatives (WO 2004/050087), aryl isoxazole derivatives (WO 2004/052087), and the like, as well as 8-heteroaryl-6-phenylimidazo [ 2004/072051) and benzophenone derivatives (WO 2005/00778) have been reported as HSP90 inhibitors useful for treating tumors.
Most of the currently known HSP90 inhibitors inhibit ATPase activity by binding to the ATP site of the N-terminal domain of HSP90, preventing the co-chaperone build-up and allowing the tumor-associated client protein to function as a proteasome ). ≪ / RTI > The attractive reason for the development of HSP90 inhibitors for clinical use is the simultaneous inhibition of various tumors and their inducing proteins, resulting in a strong antitumor effect and selectivity for cancer and normal cells, thus achieving therapeutic benefits It is.
As cited herein, 5- (2,4-dihydroxy-5-isopropylphenyl) -ene-ethyl-4- (5-methyl-1,2,4- is a compound of formula II that inhibits HSP90 activity. Oxadiazol-3-yl) isoxazole-3-carboxamide is described in WO 2011/102660.
The compound of formula (II) shows good activity in in vitro tests, but is almost insoluble in water, thereby decreasing bioavailability, and extremely low pharmacokinetic parameters have many problems in drug development.
However, the compound of formula (I) has been shown to meet all requirements as a good prodrug. In particular, it improves superior solubility and excellent pharmacokinetic properties compared to the parent compound (II).
The present invention relates to 5- (2,4-dihydroxy-5-isopropylphenyl) -ene-ethyl-4- (5-methyl-1,2,4-oxa, a compound of formula II having an inhibitory effect on HSP90 activity. It is an object to provide prodrugs for improving the pharmacokinetic properties of diazol-3-yl) isoxazole-3-carboxamide.
In order to achieve the above object, the present invention provides a novel compound represented by the following formula (I).
<Formula I>
Where
R 1 and R 2 are both PO (OH) 2 or
Particularly preferred examples of the compound of formula I according to the present invention are as follows.
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3- Phenylene bis (dihydrogen phosphate); (I-1)
(S) -2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy- 4-isopropylphenyl 2,6-diaminohexanoate dihydrochloride; (I-2)
Sodium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3 -Phenylene diphosphate; (I-3)
2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-isopropyl Phenyl dihydrogen phosphate; (I-4)
Sodium 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-iso Propylphenyl phosphate; (I-5)
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-isopropyl Phenyl dihydrogen phosphate; (I-6)
Sodium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-iso Propylphenyl phosphate; (I-7)
Potassium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3 -Phenylene diphosphate; (I-8)
Potassium 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-iso Propylphenyl phosphate; (I-9)
Potassium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-iso Propylphenyl phosphate; (I-10)
Calcium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3 -Phenylene diphosphate; (I-11)
Calcium 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-iso Propylphenyl phosphate; (I-12)
Calcium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-iso Propylphenyl phosphate; (I-13)
Magnesium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3 -Phenylene diphosphate; (I-14)
Magnesium 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-iso Propylphenyl phosphate; (I-15)
Magnesium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-iso Propylphenyl phosphate; (I-16)
The present invention also provides a process for preparing the compound of formula (I).
The preparation method of the compound of formula I according to the present invention is shown in Scheme 1 below.
<Reaction Scheme 1>
The compounds of formula (I) of the present invention may be prepared by a series of processes from the compound of formula (II), the parent drug, as shown in Scheme 1 above.
The manufacturing method will be described in more detail as follows.
1) Preparation of Compound of Formula II
Formula II compounds used as starting materials may be prepared by methods known from WO 2011/102660.
2) Preparation of Compound of Formula (I)
First, an intermediate compound having introduced protected lysine was esterified by esterification of the compound of formula II with methylene chloride in position 4 with Boc (tert-butoxycarbonyl), and then deprotected with hydrochloric acid to give R 1 or R 2
<Formula Ia>
Where
R 1 and R 2 are differently selected hydrogen or
The following scheme shows an example according to the preparation method.
Second, a bis (tetramethylphosphorodiamidate) intermediate compound is synthesized by substituting the compound of formula (II) with tetramethylphosphorodiamid chloride in a water or dioxane solvent, followed by hydrolysis using trifluoroacetic acid or hydrochloric acid. The decomposition reaction may produce the following general formula (Ib) compound wherein R 1 and / or R 2 is PO (OH) 2 .
<Formula Ib>
Where
R 1 and R 2 are both PO (OH) 2 or hydrogen or PO (OH) 2 selected differently.
The following scheme shows one embodiment of the preparation method.
Third, the compound of Formula Ib is reacted with an inorganic base selected from NaOH, KOH, Ca (OH) 2 or Mg (OH) 2 to give R 1 and / or R 2
<Formula Ic>
Where
R 1 and R 2 are both
The following scheme shows one embodiment of the preparation method.
As mentioned above, the compounds of formula I according to the invention can be used as prodrugs of compounds of formula II, which have valuable pharmacological properties.
These compounds were then investigated according to the tests given in the present invention. Evidence that the compound of formula I can be used as a prodrug of a compound of formula II, which is the parent compound, is presented according to the description given herein below.
The compound of formula (I) of the present invention has a solubility of 1,000 to 20,000 times or more compared to the compound of formula (II), which is a parent compound, and thus can increase bioavailability, and AUC 0 -120 min is 3 to 5 times and C max is greater than that of formula (II). It can be absorbed in vivo at a concentration of 10-20 folds and greatly improve pharmacokinetic properties.
Formula II compounds, which are parent compounds, are diseases that respond to inhibition of HSP90 activity, i.e. immunosuppression or inflammatory diseases such as rheumatoid arthritis, asthma, multiple sclerosis, type 1 diabetes, lupus, psoriasis and inflammation and enteropathies, viral diseases; Diabetic retinopathy, angioma and endometriosis; Or protection of normal cells against chemotherapy-induced toxicity; Hypoxia due to elevation of HSP70 in the heart and brain - diseases in which HSP90 activity is implicated, including those used for protection from ischemic injury; It is suitable for treating Scrapie / CJD, Huntington or Alzheimer's disease. Especially for the treatment of cancer.
Therefore, the compound of formula (I) according to the present invention is a prodrug in which several functional groups are introduced to the resorcinol group of the compound of formula (II), which is a parent compound, and shows effective anticancer activity against various carcinomas.
The anticancer pharmaceutical composition of the present invention may be administered by any convenient route, for example, oral, parenteral, oral, sublingual, nasal, rectal or subcutaneous, including the above formula (I) compound and a pharmaceutically acceptable excipient. It can be formulated.
In addition, the dosage or dosage of the pharmaceutical composition according to the present invention varies depending on the weight, age, sex, health condition, diet, time of administration, administration method, excretion rate and severity of the patient, the specific configuration The dose of the drug may be divided into one or several times 0.1 to 1,000 mg of the compound of formula I on an adult basis.
The present invention provides novel compounds of formula (I). The compounds of formula (I) according to the invention are used as prodrugs of compounds of formula (II) which are parent compounds, which have valuable pharmacological properties.
The compound of formula (I) of the present invention has a solubility of 1,000 to 20,000 times or more compared to the compound of formula (II), which is a parent compound, and thus can increase bioavailability, and AUC 0 -120 min is 3 to 5 times and C max is greater than that of formula (II). It is absorbed in vivo at a concentration of 10 to 20 times, which greatly improves pharmacokinetic properties and can be used to treat various tumors, including ovarian cancer and gastric cancer.
The present invention can be described in more detail through the following examples and experimental examples, but these are merely intended to illustrate the present invention, but the present invention is not limited thereto.
<Example 1> Synthesis of 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl -1,3 -phenylene Preparation of Bis (Dihydrogen Phosphate) (I-1)
Step 1: 4- (3- ( ethylcarbamoyl ) -4- (5- methyl -1,2,4 -oxadiazol- 3 - yl) isoxazol- 5 - yl) -6 -isopropyl- 1 , 3 -phenylene Preparation of bis (tetramethyl-dimethyl phosphorothioate amidate)
5- (2,4-Dihydroxy-5-isopropylphenyl) -N-ethyl-4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazole-3-car Copyamide (250 mg, 0.67 mmol) was dissolved in methylene chloride (7 mL), and then tetramethylphosphorodiamid chloride (0.35 mL, 2.35 mmol), 4- (dimethylamino) pyridine (41 mg, 0.34 mmol) was added sequentially. 1,8-diazabicyclo [5.4.0] -undec-7-ene (0.25 mL, 1.68 mmol) was added dropwise, and the reaction solution was stirred overnight at room temperature. The reaction was terminated with saturated aqueous ammonium chloride solution and extracted with methylene chloride. The organic layer was washed with saturated aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The residue was concentrated under reduced pressure, and the obtained residue was separated by column chromatography to obtain the intermediate compound 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) eye. Soxazol-5-yl) -6-isopropyl-1,3-phenylene bis (tetramethylphosphorodiamidate) (428 mg, 0.67 mmol, 99%) was obtained.
1 H-NMR (400 MHz, CDCl 3 ) δ 7.54 (s, 1H), 7.44 (s, 1H), 6.97 (br t, 1H), 3.49 (m, 2H), 3.27 (sept, 1H), 2.80 ( s, 6H), 2.77 (s, 6H), 2.66 (s, 6H), 2.63 (s, 6H), 2.61 (s, 3H), 1.26 (t, 3H), 1.19 (d, 6H)
Step 2: 4- (3- ( ethylcarbamoyl ) -4- (5- methyl -1,2,4 -oxadiazol- 3 - yl) isoxazol- 5 - yl) -6 -isopropyl- 1 , 3 -phenylene Preparation of Bis (Dihydrogen Phosphate)
The intermediate compound (428 mg, 0.67 mmol) prepared in step 1 was dissolved in a mixed solvent of trifluoroacetic acid (1.8 mL) and water (0.2 mL), and then stirred at room temperature overnight. All solvents were removed under reduced pressure, and the obtained residue was diluted with methanol. The insoluble material produced at this time was removed by filtration, and the filtrate was concentrated under reduced pressure. The obtained residue was diluted with water, washed with ethyl acetate and the aqueous layer was concentrated under reduced pressure. The material thus obtained was dissolved in water and passed through a Dowex® 50WX4 cation exchange resin to remove by-products and dried in vacuo to afford the target compound (250 mg, 0.47 mmol, 70%).
1 H-NMR (400 MHz, D 2 O) δ 7.49 (s, 1H), 7.22 (s, 1H), 3.35 (q, 2H), 3.24 (sept, 1H), 2.50 (s, 3H), 1.14 ( t, 3H), 1.11 (d, 6H)
<Example 2> (S) -2- (3- ( ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) - 5 -hydroxy -4 -isopropylphenyl 2,6 -diaminohexanoate Preparation of Dihydrochloride (I-2)
Step 1: (S) -2- (3- ( ethylcarbamoyl ) -4- (5- methyl -1,2,4 -oxadiazol- 3 - yl) isoxazol- 5 - yl) -5- hydroxy-4-isopropyl-phenyl-2,6-bis((tert-butoxy carbonyl) amino) hexanoate Preparation of
5- (2,4-Dihydroxy-5-isopropylphenyl) -N-ethyl-4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazole-3-car Copyamide (138 mg, 0.37 mmol) was dissolved in DMF (3.0 mL) to dissolve 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (248 mg, 1.29 mmol) and 4- (dimethylamino) pyridine. (22 mg, 0.19 mmol) and Boc-Lys (Boc) -OH (384 mg, 1.11 mmol) were added dropwise, followed by stirring at room temperature for 12 hours. The residue obtained by concentrating the reaction solution was separated by column chromatography to obtain an intermediate compound (S) -2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazole-3- I) isoxazol-5-yl) -5-hydroxy-4-isopropylphenyl 2,6-bis ((tert-butoxycarbonyl) amino) hexanoate (200 mg, 0.19 mmol, 53%) Got.
1 H-NMR (400 MHz, CDCl 3 ) δ 7.65 (s, 1H), 7.43 (s, 1H), 5.21 (br, 2H), 4.55 (br, 2H), 4.40 (br, 2H), 3.48 (m , 2H), 3.18-3.06 (m, 3H), 2.67 (s, 3H), 1.98 (br, 2H), 1.84 (br, 1H), 1.48 (m, 18H), 1.28-1.23 (m, 9H)
Step 2: (S) -2- (3- ( ethylcarbamoyl ) -4- (5- methyl -1,2,4 -oxadiazol- 3 - yl) isoxazol- 5 - yl) -5- Hydroxy -4 -isopropylphenyl 2,6 -diaminohexanoate Preparation of dihydrochloride
The intermediate compound (50 mg, 0.05 mmol) prepared in step 1 was dissolved in 4.0M dioxane solution (0.4 mL, 1.46 mmol) in hydrochloric acid in CH 3 CN (0.5 mL), and stirred at room temperature for 12 hours. The reaction solution was concentrated to obtain the target compound (28 mg, 0.05 mmol, 99%).
1 H-NMR (400 MHz, D 2 O) δ 7.85 (s, 1H), 7.58 (s, 1H), 4.55 (m, 1H), 4.31 (m, 1H), 3.44 (m, 2H), 3.41 ( m, 3H), 3.13 (m, 1H), 3.04-2.95 (m, 5H), 2.61 (s, 3H), 1.82 (m, 2H), 1.73 (m, 1H), 1.28-1.21 (m, 9H)
<Example 3> sodium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl Propyl -1,3 -phenylene Preparation of Diphosphate (I-3)
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3- Phenylene bis (dihydrogen phosphate) (4.90g, 9.20mmol) was dissolved in water (350ml) and 1N NaOH (36.59ml, 36.59mmol) was added dropwise, followed by stirring at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the target compound (5.63 g, 9.08 mmol, 99%).
1 H-NMR (400 MHz, D 2 O) δ 7.40 (s, 1H), 7.33 (s, 1H), 3.46 (q, 2H), 3.38 (m, 1H), 2.63 (s, 3H), 1.26 ( t, 3H), 1.16 (d, 6H)
<Example 4> 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy- -4 -isopropylphenyl Dihydrogen Preparation of Phosphate (I-4)
Step 1: 5- (4- ( ethoxymethoxy ) -2 -hydroxy -5 -isopropylphenyl ) -N-ethyl-4- (5- methyl -1,2,4 -oxadiazole- 3- I) Preparation of Isoxazole - 3 -carboxamide
5- (2,4-Dihydroxy-5-isopropylphenyl) -N-ethyl-4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazole-3-car Radiantamide (5.00 g, 13.4 mmol) was dissolved in acetone (300 mL), and then added dropwise to triethylamine (2.25 mL, 16.1 mmol) and ethoxymethyl chloride (1.31 mL, 14.1 mmol) at 0 ° C. overnight. Stirred. Concentrated under reduced pressure to remove the solvent, extracted with ethyl acetate and saturated aqueous sodium chloride solution, the organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to remove the residue by column chromatography to obtain the intermediate compound 5- (2- (ethoxymeth) Methoxy) -4-hydroxy-5-isopropylphenyl) -N-ethyl-4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazole-3-carboxamide ( 1.40 g, 3.26 mmol, 24.3%).
1 H-NMR (400 MHz, CDCl 3 ) δ 7.44 (s, 1H), 6.99 (brt, 1H), 6.67 (s, 1H), 5.87 (s, 1H), 4.90 (s, 2H), 3.54-3.45 (m, 4H), 3.10 (sept, 1H), 2.60 (s, 3H), 1.38-1.21 (m, 12H).
Step 2: 5- ( ethoxymethoxy ) -2- (3- ( ethylcarbamoyl ) -4- (5- methyl -1,2,4 -oxadiazol- 3-yl) isoxazole- 5 Yl) -4 -isopropylphenyltetramethyl Preparation of Phosphorodiamidates
The intermediate compound (1.40 g, 3.26 mmol) prepared in step 1 was dissolved in methylene chloride (40 mL), and then 4- (dimethylamino) pyridine (199 mg, 1.63 mmol) and 1,8-diazaby Cyclo [5.4.0] -undec-7-ene (0.56 mL, 3.91 mmol) and tetramethylphosphorodiamid chloride (0.56 mL, 3.91 mmol) were added dropwise in order, and the reaction solution was stirred overnight at room temperature. The reaction was terminated with saturated aqueous ammonium chloride solution and extracted with methylene chloride. The organic layer was washed with saturated aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The residue was concentrated under reduced pressure, and the obtained residue was separated by column chromatography to obtain the intermediate compound 5- (ethoxymethoxy) -2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4- Oxadiazol-3-yl) isoxazol-5-yl) -4-isopropylphenyltetramethyl phosphorodiamidate (1.82 g, 3.23 mmol, 99%) was obtained.
1 H-NMR (400 MHz, CDCl 3 ) δ 7.52 (s, 1H), 7.23 (s, 1H), 6.98 (brt, 1H), 4.93 (s, 2H), 3.52-3.43 (m, 4H), 3.26 (sept, 1H), 2.77 (s, 6H), 2.74 (s, 6H), 2.60 (s, 3H), 1.27-1.22 (m, 9H), 1.18 (t, 3H)
Step 3: 2- (3- ( ethylcarbamoyl ) -4- (5- methyl -1,2,4 -oxadiazol- 3 - yl) isoxazol- 5 - yl) -5 -hydroxy- 4 - isopropyl phenyl Dihydrogen Preparation of Phosphate
The intermediate compound (1.82g, 3.23mmol) prepared in step 2 was dissolved in 1,4-dioxane (100ml), 12N hydrochloric acid (100ml) was added dropwise and stirred at room temperature for 3 hours. Concentration under reduced pressure removed the solvent and hydrochloric acid, and the obtained residue was separated by reverse phase chromatography to obtain the target compound (1.10 g, 2.42 mmol, 75%).
1 H-NMR (400 MHz, D 2 O) δ 7.33 (s, 1H), 6.79 (s, 1H), 3.30 (q, 2H), 3.07 (sept, 1H), 2.47 (s, 3H), 1.10 ( t, 3H), 1.04 (d, 6H)
<Example 5> sodium 2 (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy Preparation of Roxy -4 - isoisophenyl Phosphate (I-5)
2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-isopropyl Phenyl dihydrogen phosphate (60 mg, 0.13 mmol) was dissolved in water (4.8 mL), 1N NaOH (0.26 mL, 0.26 mmol) was added dropwise, followed by stirring at room temperature for 30 minutes. Concentration under reduced pressure removed the solvent to obtain the target compound (66 mg, 0.13 mmol, 99%).
1 H-NMR (400 MHz, D 2 O) δ 7.22 (s, 1H), 6.96 (s, 1H), 3.33 (q, 2H), 3.06 (sept, 1H), 2.49 (s, 3H), 1.12 ( t, 3H), 1.02 (d, 6H)
<Example 6> Synthesis of 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy- -2 -isopropylphenyl Dihydrogen Preparation of Phosphate (I-6)
Step 1: 5- (2- ( ethoxymethoxy ) -4 -hydroxy -5 -isopropylphenyl ) -N-ethyl-4- (5- methyl -1,2,4 -oxadiazole- 3- I) Preparation of Isoxazole - 3 -carboxamide
5- (2,4-Dihydroxy-5-isopropylphenyl) -N-ethyl-4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazole-3-car Radiantamide (5.00 g, 13.4 mmol) was dissolved in acetone (300 mL), and then added dropwise to cesium carbonate (5.25 g, 16.1 mmol), ethoxymethyl chloride (1.31 mL, 14.1 mmol) at 0 ° C., and then stirred overnight. It was. Concentrated under reduced pressure to remove the solvent, extracted with ethyl acetate and saturated aqueous sodium chloride solution, the organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to remove the residue by column chromatography to obtain the intermediate compound 5- (2- (ethoxymeth) Methoxy) -4-hydroxy-5-isopropylphenyl) -N-ethyl-4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazole-3-carboxamide ( 3.02 g, 7.01 mmol, 52.3%).
1 H-NMR (400 MHz, CDCl 3 ) δ 7.42 (s, 1H), 6.64 (s, 1H), 5.85 (s, 1H), 4.89 (s, 2H), 3.52-3.44 (m, 4H), 3.15 ~ 3.09 (m, 1H), 2.60 (s, 3H), 1.64 (s, 3H), 1.35-1.05 (m, 16H), 0.90-0.82 (m, 2H).
Step 2: 5- ( ethoxymethoxy ) -4- (3- ( ethylcarbamoyl ) -4- (5- methyl -1,2,4 -oxadiazol- 3-yl) isoxazole- 5 Yl) -2 -isopropylphenyltetramethyl Preparation of Phosphorodiamidates
The intermediate compound (3.02 g, 7.01 mmol) prepared in step 1 was dissolved in methylene chloride (80 mL), and then 4- (dimethylamino) pyridine (428 mg, 3.50 mmol) and 1,8-diazabile were added thereto. Cyclo [5.4.0] -undec-7-ene (1.21 mL, 8.41 mmol) and tetramethylphosphorodiamid chloride (1.21 mL, 8.41 mmol) were added dropwise in order, followed by stirring at room temperature overnight. The reaction was terminated with saturated aqueous ammonium chloride solution and extracted with methylene chloride. The organic layer was washed with saturated aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The residue was concentrated under reduced pressure, and the obtained residue was separated by column chromatography to obtain the intermediate compound 5- (ethoxymethoxy) -4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4- Oxadiazol-3-yl) isoxazol-5-yl) -2-isopropylphenyltetramethyl phosphorodiamidate (3.95 g, 6.99 mmol, 99%) was obtained.
1 H-NMR (400 MHz, CDCl 3 ) δ 7.50 (s, 1H), 7.24 (s, 1H), 6.96 (brt, 1H), 4.95 (s, 2H), 3.52-3.44 (m, 4H), 3.26 (sept, 1H), 2.78 (s, 6H), 2.74 (s, 6H), 2.60 (s, 3H), 1.27-1.22 (m, 9H), 1.17 (t, 3H)
Step 3: 4- (3- ( ethylcarbamoyl ) -4- (5- methyl -1,2,4 -oxadiazol- 3 - yl) isoxazol- 5 - yl) -5 -hydroxy- 2 Oh isopropyl page carbonyl die hydrogen Preparation of Phosphate
The intermediate compound (3.95 g, 6.99 mmol) prepared in step 2 was dissolved in 1,4-dioxane (200 mL), and 12N hydrochloric acid (200 mL) was added dropwise, followed by stirring at room temperature for 3 hours. Concentration under reduced pressure removed the solvent and hydrochloric acid, and the obtained residue was separated by reverse phase chromatography to obtain the target compound (2.53 g, 5.59 mmol, 80%).
1 H-NMR (400 MHz, D 2 O) δ 7.30 (s, 1H), 6.76 (s, 1H), 3.26 (q, 2H), 3.08 (sept, 1H), 2.46 (s, 3H), 1.08 ( t, 3H), 1.00 (d, 6H)
<Example 7> sodium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy Preparation of Roxy -2 - isoisophenyl Phosphate (I-7)
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-isopropyl Phenyl dihydrogen phosphate (177 mg, 0.39 mmol) was dissolved in water (3.91 mL), 1N NaOH (0.78 mL, 0.78 mmol) was added dropwise, followed by stirring at room temperature for 30 minutes. Concentration under reduced pressure to remove the solvent, the residue obtained was dissolved in water and freeze-dried to obtain the target compound (196 mg, 0.39 mmol, 99%).
1 H-NMR (400 MHz, D 2 O) δ 7.27 (s, 1H), 6.87 (s, 1H), 3.27 (q, 2H), 3.14 (sept, 1H), 2.47 (s, 3H), 1.07 ( t, 3H), 1.01 (d, 6H)
<Example 8> Potassium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl Propyl -1,3 -phenylene Preparation of Diphosphate (I-8)
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3- Phenylene bis (dihydrogen phosphate) (1.00 g, 1.87 mmol) was dissolved in water (70 mL), 1N KOH (7.46 mL, 7.46 mmol) was added dropwise, and stirred at room temperature for 30 minutes. The reaction solution was concentrated to obtain the target compound (1.11 g, 1.63 mmol, 87%).
1 H-NMR (400 MHz, D 2 O) δ 7.40 (s, 1H), 7.33 (s, 1H), 3.46 (q, 2H), 3.38 (m, 1H), 2.63 (s, 3H), 1.26 ( t, 3H), 1.16 (d, 6H)
<Example 9> potassium 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy Preparation of Roxy -4 - isoisophenyl Phosphate (I-9)
2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-isopropyl Phenyl dihydrogen phosphate (60 mg, 0.13 mmol) was dissolved in water (4.8 mL), 1N KOH (0.26 mL, 0.26 mmol) was added dropwise, followed by stirring at room temperature for 30 minutes. Concentration under reduced pressure removed the solvent to obtain the target compound (64 mg, 0.12 mmol, 94%).
1 H-NMR (400 MHz, D 2 O) δ 7.22 (s, 1H), 6.96 (s, 1H), 3.33 (q, 2H), 3.06 (sept, 1H), 2.49 (s, 3H), 1.12 ( t, 3H), 1.02 (d, 6H)
<Example 10> Potassium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy Roxy -2 -isopropylphenyl Preparation of Phosphate (I-10)
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-isopropyl Phenyl dihydrogen phosphate (150 mg, 0.33 mmol) was dissolved in water (3.31 mL), 1N KOH (0.66 mL, 0.66 mmol) was added dropwise, and stirred at room temperature for 30 minutes. After concentration under reduced pressure to remove the solvent, the obtained residue was dissolved in water and freeze-dried to obtain the target compound (162 mg, 0.30 mmol, 93%).
1 H-NMR (400 MHz, D 2 O) δ 7.27 (s, 1H), 6.87 (s, 1H), 3.27 (q, 2H), 3.14 (sept, 1H), 2.47 (s, 3H), 1.07 ( t, 3H), 1.01 (d, 6H)
<Example 11> calcium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl Propyl -1,3 -phenylene Preparation of Diphosphate (I-11)
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3- Phenylene bis (dihydrogen phosphate) (1.00 g, 1.87 mmol) was dissolved in water (70 mL), and 1N Ca (OH) 2 (3.73 mL, 3.73 mmol) was added dropwise, followed by stirring at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the target compound (1.01 g, 1.66 mmol, 89%).
1 H-NMR (400 MHz, D 2 O) δ 7.40 (s, 1H), 7.33 (s, 1H), 3.46 (q, 2H), 3.38 (m, 1H), 2.63 (s, 3H), 1.26 ( t, 3H), 1.16 (d, 6H)
<Example 12> Calcium 2 - (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy Roxy -4 -isopropylphenyl Preparation of Phosphate (I-12)
2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-isopropyl Phenyl dihydrogen phosphate (60 mg, 0.13 mmol) was dissolved in water (4.8 mL), 1N Ca (OH) 2 (0.13 mL, 0.13 mmol) was added dropwise, and stirred at room temperature for 30 minutes. Concentration under reduced pressure removed the solvent to obtain the target compound (52 mg, 0.11 mmol, 82%).
1 H-NMR (400 MHz, D 2 O) δ 7.22 (s, 1H), 6.96 (s, 1H), 3.33 (q, 2H), 3.06 (sept, 1H), 2.49 (s, 3H), 1.12 ( t, 3H), 1.02 (d, 6H)
<Example 13> calcium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy Roxy -2 -isopropylphenyl Preparation of Phosphate (I-13)
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-isopropyl Phenyl dihydrogen phosphate (100 mg, 0.22 mmol) was dissolved in water (2.2 mL), 1N Ca (OH) 2 (0.22 mL, 0.22 mmol) was added dropwise, and stirred at room temperature for 30 minutes. Concentration under reduced pressure to remove the solvent, the residue obtained was dissolved in water and freeze-dried to obtain the target compound (96 mg, 0.19 mmol, 89%).
1 H-NMR (400 MHz, D 2 O) δ 7.27 (s, 1H), 6.87 (s, 1H), 3.27 (q, 2H), 3.14 (sept, 1H), 2.47 (s, 3H), 1.07 ( t, 3H), 1.01 (d, 6H)
<Example 14> magnesium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl Propyl -1,3 -phenylene Preparation of Diphosphate (I-14)
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3- Phenylene bis (dihydrogen phosphate) (100 mg, 0.187 mmol) was dissolved in water (7 mL), and 1 N Mg (OH) 2 (0.37 mL, 0.37 mmol) was added dropwise, followed by stirring at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the target compound (79.8 mg, 0.14 mmol, 74%).
1 H-NMR (400 MHz, D 2 O) δ 7.40 (s, 1H), 7.33 (s, 1H), 3.46 (q, 2H), 3.38 (m, 1H), 2.63 (s, 3H), 1.26 ( t, 3H), 1.16 (d, 6H)
<Example 15> magnesium 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy Roxy -4 -isopropylphenyl Preparation of Phosphate (I-15)
2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-isopropyl Phenyl dihydrogen phosphate (40 mg, 0.087 mmol) was dissolved in water (3.2 mL), 1 N Mg (OH) 2 (0.087 mL, 0.087 mmol) was added dropwise, followed by stirring at room temperature for 30 minutes. Concentration under reduced pressure removed the solvent to obtain the target compound (28.5 mg, 0.06 mmol, 82%).
1 H-NMR (400 MHz, D 2 O) δ 7.22 (s, 1H), 6.96 (s, 1H), 3.33 (q, 2H), 3.06 (sept, 1H), 2.49 (s, 3H), 1.12 ( t, 3H), 1.02 (d, 6H)
<Example 16> magnesium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy Roxy -2 -isopropylphenyl Preparation of Phosphate (I-16)
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-isopropyl Phenyl dihydrogen phosphate (62 mg, 0.14 mmol) was dissolved in water (1.36 mL), 1N Mg (OH) 2 (0.14 mL, 0.14 mmol) was added dropwise, and stirred at room temperature for 30 minutes. The mixture was concentrated under reduced pressure to remove the solvent, and then the obtained residue was dissolved in water and freeze dried to obtain the target compound (42 mg, 0.11 mmol, 81%).
1 H-NMR (400 MHz, D 2 O) δ 7.27 (s, 1H), 6.87 (s, 1H), 3.27 (q, 2H), 3.14 (sept, 1H), 2.47 (s, 3H), 1.07 ( t, 3H), 1.01 (d, 6H)
< Experimental Example 1> Solubility test
In order to determine whether the compound of formula (I) of the present invention has better solubility than the parent compound (II), the solubility was measured and shown in Table 1.
The compound of formula (I) and the parent compound of formula (II) according to the present invention were put into water, respectively, and shaken vigorously for 30 seconds every 5 minutes at 20 ± 5 ° C. to confirm the degree of melting within 30 minutes.
As can be seen in Table 1, the amount of water required to dissolve 1 g of the compound of formula II, the parent compound, is 10,000 ml or more. According to the KP solubility evaluation, the compound of formula II is a poorly soluble drug that is hardly soluble in water. The compound of formula (I) according to the invention exhibited a solubility in water of 1,000 to 20,000 times higher than that of the compound of formula (II), so that the solubility in water was very good solubility of the compound of formula (I) .
Accordingly, the compound of formula (I) according to the present invention is expected to exhibit a high bioavailability as well as greatly improved physicochemical properties to be easily formulated by showing a very good solubility compared to the formula (II) compound of the parent compound.
< Experimental Example 2> Pharmacokinetic Tests in Mice
A pharmacokinetic test was conducted to determine whether the compound of formula I according to the present invention is converted into a compound of formula II (the parent drug) in vivo and whether the concentration of the compound of formula II (the parent drug) in the blood is increased. The compound of formula I according to the present invention was orally administered to Balb / c male mice at a dose of 10 ml / kg using an oral intubation tube.
Blood samples were collected at 10, 20, 30, 60, and 120 minutes after administration on the basis of time of administration and collected in a lithium heparin coated tube. The blood thus obtained was centrifuged and the supernatant (plasma) was separated and stored frozen until analysis.
Plasma samples were prepared by liquid extraction with ethyl acetate after the addition of internal standards. Quantitative analysis was carried out by LC-MS / MS method under conditions specific to the selected compound, and analysis of the compound of formula (I) and the parent compound (II) according to the present invention was carried out on test animals to which the selected compound was administered. Pharmacokinetic parameters were obtained through WinNon Lin® non-compartmental analysis software and the results are shown in Table 2.
As can be seen in Table 2, the parent compound is a compound of formula II is AUC 0 -120 min Although the area under the plasma concentration-time curve was 33,529 ng · min · l −1 , and the C max (the highest concentration in the blood) was 420 ng / ml, the compound of formula I according to the present invention The compound of formula I according to the present invention is absorbed in vivo at a concentration of 3 to 5 times AUC 0 -120 min and 10 to 20 times C max compared to the dosage and molecular weight It can be seen that the pharmacokinetic properties are greatly improved by showing very good pharmacokinetics compared to the compound of formula II.
Therefore, the compounds of formula I according to the present invention can be usefully used as anticancer pharmaceutical compositions as prodrugs of compounds of formula II.
< Experimental Example 3> In vivo anticancer activity measurement test
In order to evaluate the anticancer efficacy of the compound of formula I according to the present invention in vivo, the following test was carried out.
A2780 ovarian cancer cell line was cultured at 8 x 10 < 6 > concentration Balb / c After xenograft subcutaneously into female nude mice, mice with tumors of appropriate size (50-200 mm 3) were selected as test animals (n = 3-6 / test group). Formula I compounds (I-1, I-2, I-3, I-7) according to the present invention were dissolved in sterile materials and administered orally at a dose of 200 mg / kg, respectively. Only the base itself was orally administered to the control group. The dosing schedule was five consecutive days (Monday to Friday) once a day for two weeks.
Body weight and tumor size of the test animals was twice measured a week, especially in tumor size is a measurement of the long axis (l) and the speed (w) of the tumor with a digital caliper, and [l × w 2] / 2 (㎣) Calculated by the formula. After calculating the average tumor size of each group, the size change value of the test group was divided by the control group size change value, multiplied by 100 and subtracted from 100% to obtain the tumor growth inhibition rate (% TGI) is shown in Table 3. Expressed as an expression:
Tumor growth inhibition rate (% TGI) = (1- drug treatment / control) x 100
As can be seen in Table 3, the compounds according to the invention showed a high tumor growth inhibition rate (% TGI).
Therefore, from the above Experimental Examples 1, 2 and 3, the new compound of the general formula (I) according to the present invention has solubility of 1,000 to 20,000 times or more than the parent compound of the general formula (II), and AUC 0 compared to the dosage and molecular weight of the compound of the general formula (II) . It can be seen that it can be very useful as an anticancer pharmaceutical composition by absorbing in vivo at a concentration of -120 min increased 3 to 5 times and C max 10 to 20 times.
Claims (11)
<Formula I>
In this formula,
R 1 and R 2 are both PO (OH) 2 or
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3- Phenylene bis (dihydrogen phosphate);
(S) -2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy- 4-isopropylphenyl 2,6-diaminohexanoate dihydrochloride;
Sodium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3 -Phenylene diphosphate;
2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-isopropyl Phenyl dihydrogen phosphate;
Sodium 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-iso Propylphenyl phosphate;
4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-isopropyl Phenyl dihydrogen phosphate;
Sodium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-iso Propylphenyl phosphate;
Potassium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3 -Phenylene diphosphate;
Potassium 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-iso Propylphenyl phosphate;
Potassium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-iso Propylphenyl phosphate;
Calcium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3 -Phenylene diphosphate;
Calcium 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-iso Propylphenyl phosphate;
Calcium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-iso Propylphenyl phosphate;
Magnesium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -6-isopropyl-1,3 -Phenylene diphosphate;
Magnesium 2- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-4-iso Propylphenyl phosphate;
Magnesium 4- (3- (ethylcarbamoyl) -4- (5-methyl-1,2,4-oxadiazol-3-yl) isoxazol-5-yl) -5-hydroxy-2-iso Propylphenyl Phosphate
<Formula II>
<Formula II>
<Formula II>
<Formula Ib>
In this formula,
R 1 and R 2 are both PO (OH) 2 or hydrogen or PO (OH) 2 selected differently.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/KR2012/006043 WO2013015661A2 (en) | 2011-07-28 | 2012-07-27 | Novel prodrugs of 5-(2,4-dihydroxy-5-isopropylphenyl)-n-ethyl-4-(5-methyl1-1,2,4-oxadiazol-3-yl)isoxazole-3-carboxamide |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR20110074867 | 2011-07-28 | ||
| KR1020110074867 | 2011-07-28 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR20130018548A true KR20130018548A (en) | 2013-02-25 |
Family
ID=47897259
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020120082521A KR20130018548A (en) | 2011-07-28 | 2012-07-27 | Novel prodrugs of 5-(2,4-dihydroxy-5-isopropylphenyl)-n-ethyl-4-(5-methyl-1,2,4-oxadiazol-3-yl)isoxazole-3-carboxamide |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR20130018548A (en) |
-
2012
- 2012-07-27 KR KR1020120082521A patent/KR20130018548A/en not_active Application Discontinuation
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20230105745A1 (en) | Cycloalkyl and hetero-cycloalkyl inhibitors, preparation methods therefor, and use thereof | |
| JP4053073B2 (en) | Isethionate, a selective CDK4 inhibitor | |
| ES2859510T3 (en) | Pyridazinone fused tricyclic compounds useful for treating orthomyxovirus infections | |
| AU2012298813B2 (en) | Inhibitors of Nedd8-activating enzyme | |
| AU2022269073A1 (en) | Exatecan derivatives and antibody-drug conjugates thereof | |
| MX2007013780A (en) | 2-amido-6-amino-8-oxopurine derivatives as toll-like receptor modulators for the treatment of cancer and viral infections, such as hepatitis c. | |
| AU2015330554B2 (en) | Crystal form of bisulfate of JAK inhibitor and preparation method therefor | |
| JP2019509990A (en) | 3,5-disubstituted pyrazoles useful as checkpoint kinase 1 (CHK1) inhibitors, and their preparation and use | |
| JP6501773B2 (en) | Salt of dasatinib in crystalline form | |
| CN112010839B (en) | Crystalline forms of a targeted silk/threonine kinase inhibitor | |
| JP2022521081A (en) | Amide derivatives useful in the treatment of HBV infections or HBV-induced diseases | |
| JP2021512922A (en) | Heteroaryl compounds, their pharmaceutical compositions, and their therapeutic use | |
| RU2680138C2 (en) | Tricyclic gyrase inhibitors | |
| US20130005696A1 (en) | Bufadienolide derivatives, preparing process thereof, composition comprising the same and the use thereof | |
| CA2956788A1 (en) | Fumarate analogs and uses thereof in the treatment of an autoimmune disease or an inflammatory disease | |
| RU2703454C2 (en) | Dipicolylamine derivatives and pharmaceutical use thereof | |
| KR20180085814A (en) | Preparation of Substituted 5,6-dihydro-6-phenylbenzo [f] isoquinolin-2-amine | |
| JP2018502851A (en) | Simple synthesis of urea derivatives of amphotericin B | |
| CN108794517B (en) | Arginase inhibitor and preparation method and application thereof | |
| WO2021129841A1 (en) | Compound used as ret kinase inhibitor and application thereof | |
| US10464883B2 (en) | Compounds and methods for the treatment of neurodegenerative diseases | |
| WO2020173417A1 (en) | Acryloyl-containing nuclear transport regulator and uses thereof | |
| JP7229482B2 (en) | Novel amide-based compound, and Pin1 inhibitor, therapeutic agent for inflammatory disease, and therapeutic agent for cancer using the same | |
| KR20130018548A (en) | Novel prodrugs of 5-(2,4-dihydroxy-5-isopropylphenyl)-n-ethyl-4-(5-methyl-1,2,4-oxadiazol-3-yl)isoxazole-3-carboxamide | |
| JP2023538638A (en) | Pyrazole boronic acid compounds, pharmaceutical compositions containing the same, and uses thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E902 | Notification of reason for refusal | ||
| E601 | Decision to refuse application |