JP2643426B2 - Pharmaceutical composition for parenteral administration - Google Patents

Description

The present invention relates to a parenteral composition comprising a drug which is insoluble or poorly soluble in water and / or is unstable in water, combined with a specific cyclodextrin derivative. It relates to an aqueous drug solution for administration. This aqueous solution provides a means of reducing the problems associated with drug deposition at the injection site and / or lungs and other organs after parenteral administration.

[Prior Art] Cyclodextrins are cyclic oligosaccharides. The most common cyclodextrins are α-cyclodextrin consisting of a ring of 6 glucose residues, β-cyclodextrin consisting of a ring of 7 glucose residues, and γ-cyclodextrin consisting of a ring of 8 glucose residues. Cyclodextrin. The interior cavity of the cyclodextrin is lipophilic and the exterior of the cyclodextrin is hydrophilic. Due to this combination of properties, extensive research has been conducted on natural cyclodextrins, especially in combination with drugs, and numerous inclusion compounds have been reported. Although β-cyclodextrin has received particular attention due to its internal cavity size, its relatively low water solubility has limited its use in the pharmaceutical field.

Attempts to modify the properties of natural cyclodextrin have resulted in heptakis (2,6-di-O-methyl) -β-cyclodextrin, heptakis (2,3,6-tri-O-methyl) -β-cyclodextrin, Hydroxypropyl-
Cyclodextrin derivatives such as β-cyclodextrin, β-cyclodextrin-epichlorohydrin polymer have been developed. For information on cyclodextrins and their use in research in the pharmaceutical field, see, for example, Pitha
et al, Controlled Drug Delivery (ed. by SDBruck), Volume 1, CRS Press, Florida, USA, pp. 125-148 (1983). For a more recent overview, Ue
kama et al., CRC Critical Reviews in Therapeutic Drug C
arrier Systems , Vol. 3 (1), 1-40 (1987); Uekama, To
pics in Pharmaceutical Sciences 1987 (DDBreimer
Other editions), Elsevier Science Publishers BV (Biomedica
l Division), 1987, 181-194; and Pagington, Chemist
See also ry in Britain, May 1987, 455-458.

For inclusion compounds of α-, β- or γ-cyclodextrin or mixtures thereof with various agents,
Many proposals have been made so far, and various advantages of this clathrate have been pointed out. These prior arts disclosed in US patents are summarized below.

2,6-di-O-methyl-β-cyclodextrin;
Inclusion compounds with dibenzo [bd] pyran derivatives and salts having analgesic, vomiting suppressing and narcotic potentiating effects are Ngr
di et al., US Patent No. 4,599,327, describes this inclusion compound as having increased water solubility and therefore improved biological activity. Regarding the application of such methylated cyclodextrins to pharmaceuticals,
It is described in Uekama, Pharm, Int., March 1985, 61-65. Pitha, J. of Inclusion Phenomana, 2,477-485 (198
See also 4).

For polymers of cyclodextrin, see Fenyvesi
Et al . , Chem. Pharm. Bull. 32 (2), 665-669 (1984).
It has been reported to improve the dissolution of furosemide. The use of water-soluble β-cyclodextrin epichlorohydrin polymers to improve phenytoin dissolution and absorption has been described by Uekama et al., International Journal of Pharm.
aceutics , 23 , 35-42 (1985).

Hydroxypropyl-β-cyclodextrin (HPC
The production of HPCD by the addition of propylene oxide to D) and β-cyclodextrin has been described nearly 20 years ago.
No. 3,459,731 to Gramera et al. G
Ramera et al. also reacted β-cyclodextrin with ethylene oxide in a similar manner to give hydroxyethyl-
Producing β-cyclodextrin is also reported. More recently, Pitha and coworkers have reported an improved method of making this cyclodextrin derivative and its effect on the dissolution of various drug molecules. Pitha U.S. Pat.No. 4,596, June 24, 1986,
795 discloses inclusion compounds of sex hormones, especially testosterone, progesterone and estradiol, with certain cyclodextrins, preferably hydroxypropyl-β-cyclodextrin and poly-β-cyclodextrin. This inclusion compound can successfully supply sex hormones to the general circulation via the sublingual or buccal route. The effectiveness of this supply is attributed to "the high solubility of the hydrophilic derivative of cyclodextrin, the non-aggregated structure of the inclusion compound with this steroid, and its low toxicity and low irritation of oral tissues" Can be The success of other cyclodextrins, including poly-γ-cyclodextrin and hydroxypropyl-γ-cyclodextrin, is also mentioned in the above-mentioned Pitha US patent. Pitha et al . , J. Pharm. Sci. , Vol . 74, No. 9, September 1985, for the same and related work as described above .
See also Moon, 987-990. The article in Pitha et al . In J. Pharm. Sci. Includes testosterone / hydroxypropyl- β-
The storage stability of tablets containing cyclodextrin clathrates, the lack of toxicity of cyctodextrin itself, and the amorphous nature of cyclodextrin derivatives and their clathrates with drugs are important for improving dissolution properties thing,
Are also described.

Improved optimized production and purification methods for hydroxypropyl-β-cyclodextrin have recently been described by Pitha et al., Int.
ernational Journal of Phamaceutics , 29 , 73-82 (198
6). In this document, the author states that hydroxypropyl-β-cyclodextrin has a high concentration (40
(.About.50%) states that the aqueous solubility of 32 drugs in aqueous solution was increased, including: acetamidephen, apomorphine, butylated hydroxytoluene, chlorthalidone, cholecalciferol, dexamethasone, dicumarol, digoxin, Diphenylhydantoin, estradiol, estriol, ethinyl estradiol 3-methyl ether, ethisterone, furosemide, hydroflumethiazide, indomethacin, isoproniazide phosphate, 17-methyltestosterone, nitroglycerin, norethindrone, ouabain, oxprenolol, progesterone, Improves solubilization of retinal, retinoic acid (all trans and salt forms), retinol, spironolactone, sulpiride, testosterone, and theophylline It was observed. The authors of the above references also note that this study builds on the earlier work they did on hydroxypropyl-β-cyclodextrin, which had previously been found to be effective for oral administration of sex hormones to humans. Yes, he said. Pitha
Et al., International Journal of Pharmaceutics , 29 , 73
−89 (1986), their subsequent study,
Most recently issued Ptha U.S. Patent No. 4,727,064 (19
88.2.23). This patent includes compositions comprising an amorphous inclusion compound of cyclodextrin and a drug, and (1) two or more water-soluble cyclodextrin derivatives capable of forming an inclusion compound with the drug. Dissolving the essentially amorphous mixture in water, and (2) solubilizing the lipophilic drug in an aqueous medium to form a solution, forming a solubilized drug / cyclodextrin inclusion compound. A process for the preparation of a stabilizing amorphous inclusion compound of a drug and a cyclodextrin mixture is described.

Uekama et al., CRC Critical Reviews in Therapeutic Dr
ug Carrier Systems , Vol. 3 (1), pp. 1-40 (1987) describes the properties of various cyclodextrins including hydroxypropyl-β-cyclodextrin.
The authors reported that the presence of 15 mg / ml HPCD improved the water solubility of the carmofur, diazepam, digitoxin, digoxin, flurbiprofen, indomethacin, isosorbide dinitrate, phenytoin, prednisolone, progesterone, and testosterone drugs. Posted data indicating that Regarding metabolism and toxicity of cytodextrins, Uekama et al. Show that cyclodextrins at significantly higher concentrations cause hemolysis, and that methylated cyclodextrins, in particular, have a higher hemolytic effect than native cyclodextrins. Points out. Hydroxypropyl-β-cyclodextrin is said to initiate hemolysis at 4.5 mM. The author further added:
Parenteral administration of large amounts of cyclodextrins should be avoided, but γ-cyclodextrin and hydroxypropyl-β-cyclodextrin appear to be useful for solubilizing drugs for injection and for liquid preparations for mucosal use. Points out.

JANSSEN PHARMACEUTICA NV International Patent Application No.PCT / E
P84 / 00417 (released on July 4, 1985, International Publication No. WO85 / 027
67) Pharmaceutical compositions comprising an inclusion compound of a drug which is unstable or only slightly soluble in water and a partially etherified β-cyclodextrin derivative having a hydroxyalkyl group and optionally further an alkyl group Things are listed. The cyclodextrin derivatives also include hydroxypropyl-β-cyclodextrin, while the drugs include non-steroidal anti-rheumatic drugs, steroids, cardiac glycosides, and benzodiazepines, benzimidazoles, piperidines, piperazines, imidazoles and triazoles. Derivatives are included. Preferred agents include etomidate, ketoconazole, tubrazole, itraconazole, levocabastine and flunarizine. The pharmaceutical compositions of this international application include formulations for oral, parenteral, and topical administration, and solubilization of various drugs may require 4 to 10 of cyclodextrin derivatives.
% Solution is used. Indomethacin, digitoxin, progesterone, dexamethasone, using 10% HPCD
The water solubility of hydrocortisone and diazepam has been shown to be improved, and an injection of diazepam in 7% HPCD is described as an example. The relatively low cyclodextrin concentration was used because one wanted to avoid or minimize the hemolytic effects seen at high cyclodextrin concentrations.

Carpenter et al., The Journal of Pediatrics, 111, 507-
512 (October 1987) describes the treatment of severe hypervitaminosis A by intravenous infusion of 2-hydroxypropyl-β-cyclodextrin prepared as a 5% solution in water. It was observed that circulating retinyl ester temporarily increased during the instillation, but the total amount of vitamin A excreted in the urine after the infusion increased. For example, intravenous infusion of 5% HPCD has been found to reduce the in vivo concentration of vitamin A, which probably forms an inclusion compound with this vitamin and removes some excess from the body. It is probably due to.

 Production of inclusion compounds of other cyclodextrin derivatives
The composition is also described in the literature. α-, β- and
Glucosyl and maltosyl of γ-cyclodextrin
A branched cyclodextrin derivative which is a derivative, and
Research on clathrates and their inclusion has recently been reported
ing. Uekama, IA USTopics in Pharmaceutical Sciences 1
987(D.D.Breimer et al.), Elsevier Science Publishers
 B.V. (Biomedical Division), 1987, 181-194,
Of rutosyl and glucosylcyclodextrin derivatives
Effects on biopharmaceutical properties, including increased drug absorption
Have been described. Koizumi et al.Chem.Pharm.Bull.,35
(8), 3413-3418 (1987) describes a poorly water-soluble drug and
Lucosyl cyclodextrins, ie, 6-O-α
-D-glucosyl-a-CD (G₁-Α-CD), 6-O-α
-D-glucosyl-β-CD (G₁-Β-CD) and 6^A, 6^D
-Di-O-α-D-glucosyl-β-CD (2G₁-Β-C
Inclusion compounds with D) have been reported. Okada et al.Chem.
Pharm.Bull.,36(6), 2176-2185 (1988)
Solution drugs and maltosyl cyclodextrins,
6-O-α-maltosyl-α-CD (G_Two-ΑCD), 6
-O-α-maltosyl-β-CD (G_Two-Β-CD), 6-O
-Α-maltosyl-γ-CD (G_Two-Γ-CD), 6-O-α
-Maltotriosyl-α-CD (G_Three-Α-CD), 6-O-
α-maltotriosyl-β-CD (G_Three-Β-CD) and 6
-O-α-maltotriosyl-γ-CD (G_Three-Γ-CD) and
Have been reported.

The supply of drugs to the brain is dependent on transport and metabolic factors, more specifically on the functional barrier of the endothelial brain capillary wall,
That is, it is often significantly restricted by the blood-brain barrier (BBB). Site-specific and sustained delivery of drugs to the brain is even more difficult.

It has recently been found that the application of the dihydropyridine pyridinium salt redox system to the delivery of a number of drugs to the brain has been successful. Generally described, the redox system is used to synthesize a dihydropyridine derivative of a biologically active compound. This derivative can enter the CNS (Central Nervous System) through the blood-brain barrier after its systemic administration. Thereafter, when the dihydropyridine-type compound is oxidized to the corresponding pyridium salt, the drug is delivered to the brain.

So far, the following three main methods have been announced for supplying drugs to the brain using this redox system.

The first method is to form a derivative containing a pyridinium nucleus as an integral structural part from a selected drug. In this method, N-methylpyridinium-2-carbaldoxime chloride (2-PAM) whose active nucleus constitutes a quaternary pyridium salt is converted to a latent prodrug of the dihydropyridine type (precursor drug species). It was first applied to supply the brain via morphology. That is, by making the hydrophilic compound (2-PAM) into its dihydropyridine form (Pro-2-PAM), it can be made lipoid (that is, lipophilic or lipophilic) and can pass through the lipoid barrier. I did it. Although this simple prodrug method was able to introduce this compound into the brain and other organs, this operation did not actually yield any brain specificity, nor did it It was impossible. Moreover, the application of such a method is limited to relatively small molecule quaternary pyridinium ring-containing drug species, and continuously releases the desired drug specifically to the brain, and at the same time, rapidly removes it from the general circulatory system. Did not produce the overall ideal result with increased potency and reduced toxicity. Capture of 2-PAM produced in the brain (tra
pping) did not occur, and consequently, no brain-specific sustained supply occurred. Generated 2-PAM
Was also rapidly cleared from the brain at a rate similar to that from the general circulation and other organs. In this regard, U.S. Pat. Nos. 3,929,813 and 3,962,447; Bodor et al . , J. Pharm. Sci. , 67 , No. 5,685.
(1978). In addition, Roche, EB Design
of Biopharmaceutical Properties Through Prodrugs
and Novel Approaches for the Desi
gn of Membrane Transport Properties of Drugs ", APhA Academy of Pharmaceutical
See also Sciences, Washington DC, 98-135 (1976). However, with the subsequent development of this first method,
Berberine, a much larger quaternary salt, was delivered to the brain via its dihydropyridine-type prodrug form and found to provide a site-specific, sustained delivery of the anticancer drug to the brain. Border et al., Science, Vol, 21
4, pp. 1370-1372 (December 18, 1981).

A second method of supplying a drug to the brain using the redox system is a method in which a pyridium carrier is chemically bound to a biologically active compound. Border et al., Science , Vol. 214, pp. 1370-1372 (1981, Jan. 1)
(February 18) outlines the specific and sustained supply of drugs to the brain in this manner, as shown in Scheme I below.

According to this scheme in Science , the drug [D] is coupled to the quaternary carrier [QC] ⁺ and the resulting [D-QC] ⁺
Is then chemically reduced to the lipoidal dihydro form [D-DHC]. When this [D-DHC] is administered in vivo, it is rapidly distributed throughout the body, including the brain. The dihydro form of [D-DHC] then undergoes in situ oxidation (rate constant, K ₁ ) (oxidation in vivo by the NADNADH system), ideally the inert [D-QC] ⁺ quaternary It becomes a grade salt. This quaternary salt is rapidly excreted from the general circulatory system of the body due to its ionic and hydrophilic properties, but its excretion from the brain is hindered by the blood-brain barrier (K >> K ₂ ; K ₃ >>
K ₇ ). Enzymatic cleavage of [D-QC] ⁺ thus “occluded” or “confined” in the brain results in a sustained supply of drug species [D], followed by normal excretion of this drug species (K ₅ ) , Metabolism occurs. A properly selected carrier [QC] ⁺ will also be rapidly excreted from the brain (K ₆ >> K ₂ ).
[D-QC] ⁺ is easily excreted from the general circulatory system, so that only a very small amount of drug is released to the body (K ₃ >>
K ₄ ), [D] will be released mainly into the brain (K ₄ >
K _2). Taken together, ideally, a brain-specific sustained release of the target drug species would be obtained. Specifically, Border and colleagues studied phenylethylamine as a drug model. This compound was coupled with nicotinic acid and then quaternized to give the compound of the formula

This compound was then reduced with sodium dithionite to give the corresponding compound of the formula

The in vivo test results of the above N-methyl derivatives supported the concept shown in Scheme I. Border et al. Believe that the above or similar carrier systems could be used to deliver a variety of drugs, and for the delivery of amino or hydroxyl group containing drugs, including small peptides, to the brain. It was noted that N-methylnicotinic acid esters and amides and the use of these pyridine ring substituted derivatives had been studied. Other specific examples of possible carriers were not disclosed.

For further information on this study using the Drex carrier system, see The Friday Evening Post, August 14, 1981 (Health Center Communications, University of Florida, Gainesville, Florida, USA); Chem.
ical & Engineering News , December 21, 1981, pp.24-25;
And Science News, January 2, 1982, Vol. 121, No. 1,
There is also a report on p.7. More recently, the redox carrier system has been substantially expanded with respect to possible carriers and delivered drugs. For example, International Patent Application No.P
CT / US83 / 00725 (filed May 12, 1983, International Publication No. W08
3/03968, published November 24, 1983).
Also, Bodor et al, Pharmacology and Therapeutics, Vo
I. 19, No. 3, pp. 337-386 (1983); and Border U.S. Patent No. 4,540,564 (September 10, 1985).

A third method for supplying a drug to the brain using the above-described redox system is a derivative in which a primary, secondary or tertiary amine functional group of a centrally acting amine is substituted with a dihydropyridine pyridinium salt type redox system. The method to use. This brain-specific analog of a centrally acting amine is described in International Patent Application No. PCT / US85 / 00236 (filed February 15, 1985, published September 12, 1985 as International Publication No. W085 / 03937). ing. Dihydropyridine type analogous compounds (derivatives) have the structural formula Indicated by Wherein D represents the residue of a centrally acting primary, secondary or tertiary amine; Represents a group represented by the following formulas (a) to (d).

In the above formula, the dashed line of formula (a) means that a double bond is present at either the 4- or 5-position of the dihydropyridine ring; Means that a double bond is present in any of
Is 0 or 1; n is 0, 1 or 2; p is 0,
1 or 2 with the proviso that when p is 1 or 2, each R group of formula (b) can be located on any of the two fused rings; q is 0, 1 or 2 Yes, but q
Is 1 or 2, each R group of formula (c) can be located on any of the two fused rings; each R group is halogen, C ₁ -C ₇ alkyl, C ₁ -C ₇ C ₇ alkoxy, C ₂ -C ₈ alkoxycarbonyl, C ₂ -C ₈ alkanoyloxy, C ₁ ~
C ₇ haloalkyl, C ₁ ~C ₇ ~ alkylthio, C ₁ -C ₇ alkylsulfinyl, C ₁ -C ₇ alkylsulfonyl, -CH = NO
R (R is a hydrogen or C ₁ -C ₇ alkyl group), and -CONR'R "(R 'and R" may be a different even different identical, each represent a hydrogen or C ₁ -C ₇ alkyl group A) independently selected from the group consisting of:

The above-mentioned dihydropyridine type analogous compound is in vivo,
Acts as a supply system for the corresponding biologically active quaternary salt compound. Due to its lipophilicity (lipophilicity), dihydropyridine-type analogs are distributed systemically and can easily penetrate the brain through the blood-brain barrier. Then, when oxidized in vivo and converted to a quaternary salt form,
Preferential "confinement" in the brain occurs. However, unlike the drug-carrier derivatives described in the aforementioned Border U.S. Pat.No. 4,540,564 and related literature, there is no readily metabolically cleavable bond between the drug moiety and the quaternary salt moiety. . The active species supplied to the brain is the quaternary salt form of the analog itself, rather than the first drug (stock solution) used to derive the dihydro analog.

As described above, the three main methods using the dihydropyridinepyridinium salt-type redox system for drug delivery targeted to the brain described above have their own unique properties, but are common to all methods. There is also a property to do.
Common to all methods is the introduction of a dihydropyridine-type nucleus into the drug molecule, such that the resulting dihydropyridine-containing drug derivative is derived from the drug (the drug substance) used to derive it. ), The affinity is substantially higher. This increase in lipophilicity allows the resulting derivative to easily penetrate biological membranes including the blood-brain barrier. Another property common to all methods is that the "redox" nature of the dihydropyridine type moiety allows this lipophilic dihydropyridine form of the compound to be oxidized in vivo to a hydrophilic, ionic pyridinium salt form. The consequence of this is that depending on the method used, the active agent itself or its quaternary salt precursor is confined in the brain.

The dihydropyridine pyridinium salt redox carrier and its analogous systems have been very successful in concentrating drug delivery to the brain in laboratory tests. Part of this success is, of course, due to the highly lipophilic nature of dihydropyridine-containing derivatives. This property allows penetration into the brain. But at the same time, this increased lipophilicity
There is also the disadvantage that it is practically impossible to prepare or formulate aqueous solutions of these derivatives for injection. further,
Even when the dihydropyridine-containing derivative is dissolved in an organic solvent such as dimethyl sulfoxide, the derivative tends to precipitate out of solution after injection, especially at higher concentrations and at the injection site or lung. In fact, it has been found that, even without appreciable crystal precipitation, the redox derivatives, in addition to exhibiting the desired concentration in the brain, often also exhibit undesirable high lung concentrations. As a result, the drug concentration ratio in the brain / blood is at an appropriately high level, while the drug concentration ratio in the initial lung / brain is also high. Moreover, dihydropyridine-containing derivatives are:
Even in the dry state, it is very sensitive to oxidation and the addition of water, so there are drawbacks to stability issues. These problems that must be overcome in order to completely commercialize the redox system of the dihydropyridine pyridinium salt type are also pointed out in Japanese Patent Application No. 64-37. In particular, the invention relates to problems associated with undesirably high concentrations or precipitation of the dihydropyridine pyridinium salt redox system at or near the injection site after parenteral administration and / or in the lung, as well as being insoluble or poorly soluble in water and / or water. Address the similar problems seen with parenteral administration of other unstable drugs.

[Problem to be Solved by the Invention] An object of the present invention is to determine whether the compound is insoluble or poorly soluble in water,
And / or to provide an improved aqueous solution for parenteral administration of an unstable drug in water.

Another object of the present invention is to provide a method of reducing the tendency of a lipophilic and / or water labile drug to precipitate after or near its parenteral administration at or near the injection site and / or in the lungs.

It is yet another object of the present invention to provide an improved aqueous solution for parenteral administration containing a dihydropyridine compound that is a reduced form of a redox system of the dihydropyridine pyridinium salt type for drug delivery to the brain.

Another object of the present invention is to provide a dihydropyridine pyridinium salt redox-based dihydropyridine compound for drug delivery targeted to the brain, wherein the dihydropyridine compound precipitates at or near the injection site and / or in the lung after parenteral administration. Is to provide a way to reduce

[Means for Solving the Problems] The object of the present invention is to provide a drug which is insoluble or hardly soluble in water and / or unstable in water by converting β- or γ-cyclodextrin hydroxypropyl, hydroxyethyl, glucosyl, maltosyl or maltotrio. It is achieved by an aqueous solution for parenteral administration dissolved in an aqueous solution containing about 20-50% of the sill derivative. The invention also relates to β- or γ
Lipophilic and / or lipophilic and / or a composition comprising a drug dissolved in an aqueous solution for parenteral administration containing about 20-50% of a hydroxypropyl, hydroxyethyl, glucosyl, maltosyl or maltotriosyl derivative of cyclodextrin. Alternatively, there is provided a novel method for reducing the tendency of a drug that is unstable in water to precipitate at or near the injection site and / or in the lungs and other organs after its parenteral administration. In one aspect of the invention, the drug is a biooxidative, blood-brain barrier permeable, lipoidal dihydropyridine compound that is a reduced form of a redox system of a dihydropyridinepyridinium salt for drug delivery to the brain.

[Operation] Hereinafter, the present invention will be described in detail.

As used herein, the term "lipophilic" means that the drug is lipid soluble and hydrophobic, ie, insoluble or sparingly soluble (almost insoluble) in water.

"Parenteral" means routes of administration other than through the gastrointestinal tract or lungs, as well as compositions (formulations) used for drug administration by such routes. That is, the term “parenteral” as used herein includes, for example, intramuscular, subcutaneous, intraarticular (including intra-articular bursa), and especially intravenous routes and formulations. It is. As used herein, the terms "parenteral" and "injectable" are used interchangeably.

Numerous drugs have problems associated with lack of water solubility and / or lack of stability in water. Such lipophilic and / or water-labile drugs are virtually impossible to formulate in the form of aqueous solutions for parenteral administration. As a result, such drugs are currently not available for injection, or are only available for injection in combination with undesirable organic excipients (vehicles).
Injection of such organic excipients is undesirable because it can cause systemic or local toxicity. Some examples of organic solvents commonly used as excipients include dimethylacetamide (DMA), dimethylsulfoxide (D
MSO), propylene glycol (PG), benzyl alcohol and ethanol. Examples of toxicity associated with these solvents include central nervous system depression, nystagmus, lymphocytosis, liver and kidney damage, blood disorders,
There are jaundice, weight loss, anemia, convulsions, hallucinations, mutagenic effects, cyanosis, hypotension, bronchospasm, cardiac arrest, and death.

In addition, parenteral administration of a solution of a lipophilic or water-labile drug in an organic vehicle may cause the drug to precipitate out of solution at or near the injection site and / or in the lungs and other organs. Also lead to increased toxicity. For example, drug deposition in the lungs has led to severe respiratory distress in animal studies and has sometimes been fatal. On the other hand, in the case of drugs that are only available for oral administration due to lack of a suitable solvent, the drugs are generally less bioavailable in oral administration than in parenteral forms, especially in intravenous routes. Therefore, the bioavailability of the drug becomes a problem.

The lipophilic and / or water-labile drugs that may be used in formulating the aqueous solution for parenteral administration according to the present invention include the following drugs: anti-neoplastic drugs (anticancer / anti-tumor drugs) Tumor drugs), sedatives, anti-inflammatory steroids,
Tranquilizer, anticonvulsant, antiviral, vitamin /
Nutritional factors, emetics, anticoagulants, inotropics (including cardiac glycosides), diuretics, non-steroidal anti-inflammatory drugs (NSAIDs), androgens, estrogens, vasodilators, antidepressants, hypnotics, Antifungal, Progestin, Antigenic animal, Anesthetic, Vasoconstrictor, Hypoglycemic, Antibacterial / antibiotic, Platelet inhibitor, Muscle relaxant, Antiemetic, Radioactive diagnostic, Anticonvulsant, Antiarrhythmic Drugs, carbonic anhydrase inhibitors, narcotic antagonists, narcotic drugs, narcotic / antagonistic combinations, pharmacologically active proteins (eg, peptide hormones, enzymes, antibodies and other biological products), anti-Parkinson's disease / Dopming agents and therapeutic drugs for Alzheimer's disease.

Specific examples of drugs that can be used in parenteral pharmaceutical compositions in combination with hydroxypropyl-β-cyclodextrin according to the present invention include: chlorambutyl, lomustine, melphalan, methotrexate, Hexamethylmelamine, teniposide, etoposide, semustine (methyl CCNU), fazarabine (Ara-AC), mercaptopurine, tubrazole, carmofur, carmustine, amsacrine, bruceanthin, diazicon, didemnin B, echinomycin and
Antineoplastic drugs such as PCNU; anti-inflammatory steroids such as dexamethasone, hydrocortisone and prednisolone; 1
Estrogens such as 7β-estradiol, 17a-ethynylestradiol, ethinylestradiol 3-methylether and estriol; progestins such as norethindrone, norethindrone acetate, norgestrel, ethisterone, medroxyprogesterone acetate and progesterone; anti-antigens such as phenytoin (diphenol hydantoin) Seizures; hypnotics such as pentobarbital, phenobarbital and secobarbital, barbiturates with various pharmacological actions such as anticonvulsants and sedatives; antivirals such as vidarabine; retinol (vitamin A), vitamin A acetate Vitamins / nutritional factors such as, cholecalciferol and retinal, and other fat-soluble vitamins such as vitamins E, D and K; Emetics such as morphine; diuretics such as chlorthalidone, furosemide and spironolactone; anticoagulants such as dicoumarol; inotropics such as digoxin and digitoxin; nonsteroidal anti-inflammatory drugs such as indomethacin, piroxicam and flurobiprofen; Androgens such as methyltestosterone and testosterone; steroidal hypnotics / anaesthetics such as alphaxalone; antidepressants such as sulpiride; antibiotics such as ampicillin and penicillin G; coronary vasodilators such as nitroglycerin and flunarizine;
Hypnotics such as etomidate; carbonic anhydrase inhibitors such as acetazolamide; ketoconazole, itraconazole,
Antifungal drugs such as metronidazole and miconazole benzoate; antigenic animal drugs such as flubendazole; anesthetics such as lidocaine; hypoglycemic drugs such as acetohexamide; antiemetics such as dimenhydrinate; Antibacterials; dopamine agonists such as L-dopa; THA
Benzodiazepines having various pharmacological actions such as sedatives, hypnotics, anticonvulsants, tranquilizers and muscle relaxants, such as chlordiazepoxide, diazepam, medazepam, oxazepam and lorazepam; and prostaglandin Gins, for example,
PGEs such as PGE ₁ (alprostadil), a vasodilator, and PGI ₂ (prostacyclin or epoprostenol), a platelet inhibitor.

In a particularly preferred embodiment of the present invention, an anti-neoplastic drug (anti-cancer drug) is used as a drug in the pharmaceutical composition for parenteral administration of the present invention. Particularly suitable are anti-neoplastic agents such as chlorambutyl, lomustine, melphalan, hexamethylmelamine, methotrexate, semustine, teniposide, etoposide and fazarabine.

In another preferred embodiment of the present invention, alphaxalone, which is a steroidal anesthetic / hypnotic, is used as a drug in the composition for parenteral administration of the present invention.

In still another preferred embodiment of the present invention, as the drug in the composition for parenteral administration of the present invention, a dihydropyridine derivative which is a reduced form of a dihydropyridinepyridinium salt-type redox system for drug delivery to the brain is used as the drug. Use as

For the redox system for drug delivery to the brain, the following definitions apply herein.

The term "lipoidal" refers to a redox form or site that is lipid soluble or lipophilic.

The terms "redox carrier system" and "redox-like system" refer to two different methods of centrally delivering drugs to the brain using a system of the dihydropyridinepyridinium salt type, and are used in either of these methods The compound is used in combination with the cyclodextrin derivative selected in the present invention.

In other words, the redox carrier system supplies a drug targeting the brain by a carrier / drug conjugate. This can be represented by the following formula in its reduced form, which is the form used for administration.

[D-DHC] In the above formula, [D] means a centrally acting drug,
[DHC] means a biooxidative, blood-brain barrier permeable, lipoidal reduced form (ie, dihydropyridine form) of a dihydropyridine pyridinium salt redox carrier. On the other hand, the oxidized form of the redox carrier system that is "confined" in the brain (from which the active drug is ultimately released) can be represented by the following formula:

In the above formulas, X ^{- -} "D-QC] ⁺ X denotes an anion of non-toxic acceptable acids to the drug, means [D] is a drug types centrally acting,
[QC] ⁺ means a hydrophilic, ionic pyridinium salt form of a dihydropyridine pyridinium salt type redox carrier. Various methods using the redox carrier system have already been described in the section of [Prior Art]. This redox carrier system is of the type described as the second method in the above description among redox systems developed for supplying a drug to the brain.

Various uses of the redox carrier system are described in detail in the following patent documents: Boder U.S. Pat. Nos. 4,479,932 (1984.10.30) and 4,540,564 (1985.9.10); No. 4,617,298 (198
6.10.14), and International Application No. PCT / US83 / 00725 of the University of Florida (International Publication No. W083 / 03968,1
983.11.24).

Redox-like systems also provide a brain-targeted drug delivery with a novel compound containing a dihydropyridinepyridinium salt-type moiety, but unlike redox carrier systems, are metabolically easier to release the original drug molecule. It does not cleave.

One example of a method utilizing a redox-like system is to replace the primary, secondary or tertiary amine function of a centrally acting amine with
It forms a derivative substituted by a dihydropyridine pyridinium salt type redox system, and has already been described in the section of [Prior Art]. This redox-like system belongs to the type described as the third method of the above-mentioned redox system developed for supplying drugs to the brain.
The various uses of this analogous system are the University of
Florida International Application No. PCT / US85 / 00236 (International Publication No.
85/03937, 1985.9.12).

Another method utilizing a redox-like system is to form novel amino acid and peptide derivatives containing a dihydropyridine pyridinium salt-type redox moiety, and this derivative converts the redox system to a carbon atom adjacent to the carboxyl carbon of the amino acid. It is bonded to an atom directly or via an alkylene bonding group. These amino acids and peptides are described in detail in University of Florida, JP-A-63-277672. In summary, the novel redox amino acids are compounds having the following structural formula in their reduced form.

Wherein, Z denotes either a direct bond or C ₁ -C ₆ alkylene group may be bonded through a ring carbon atom or ring nitrogen atom to the nitrogen-containing heterocyclic ring; in Z is ring carbon atoms When linked, R ₁ is C ₁ -C ₇ alkyl, C ₁ -C ₇
A haloalkyl or a C ₇ -C ₁₂ aralkyl group; when Z is attached to a ring nitrogen atom, R ¹ represents a direct bond; R ₂ and R ₃ may be the same or different and each hydrogen, halogen, cyano, C ₁ -C ₇ alkyl, C ₁ -C ₇ alkoxy, C ₂ -C ₈ alkoxycarbonyl, C ₂ -C ₈ alkanoyloxy, C ₁ -C ₇ haloalkyl, C ₁ -C ₇ alkylthio,
C ₁ -C ₇ alkylsulfinyl, C ₁ -C ₇ alkylsulfonyl, -CH = NOR (R is a hydrogen or C ₁ -C ₇ alkyl group), or -CONR'R "(R 'and R"
May be the same or different and are each hydrogen or C ₁ -C ₇
Or one of R ₂ and R ₃ together with an adjacent ring carbon atom may form a benzene ring fused to the heterocycle, wherein the benzene ring is Accordingly, hydroxy, protected hydroxy, halogen, cyano, C ₁ -C ₇ alkyl, C ₁ -C ₇ alkoxy, C ₂ -C ₈ alkoxycarbonyl, C ₂ -C ₈ alkanoyloxy, C ₁ -C ₇ haloalkyl, C ₁ ~C ₇ alkylthio,
C ₁ -C ₇ alkylsulfinyl, C ₁ -C ₇ alkylsulfonyl, -CH = NOR (R is a hydrogen or C ₁ -C ₇ alkyl group), and -CONR'R "(R 'and R"
May be the same or different and are each hydrogen or C ₁ -C ₇
R ₄ has a hydrogen or carboxyl protecting group; R ₅ has the same or different substituent (s) selected from the group consisting of: With hydrogen or amino protecting groups; and the dotted line indicates that the compound is 1,4- or 1,6-dihydropyridine, 1,4 or
It has a 1,2-dihydroquinoline or 1,2-dihydroisoquinoline ring system.

The compounds in the reduced form of the novel dihydrolysine amino acid analogs shown above and the corresponding oxidized salts thereof are useful for the production of novel redox-containing peptides having the partial structure represented by the following formula.

and The novel peptide analogs having partial structure (A) act as a supply system for the corresponding quaternary salt of partial structure (B) in vivo. This quaternary salt-type derivative is also used as a chemical intermediate for producing the above-mentioned dihydro-type compound, but is itself pharmacologically active in vivo, or it is pharmacologically active in vivo. When administered in the form of the corresponding reduced form of dihydropyridine, it provides site-specific and sustained drug delivery to the brain. The production of these amino acid analogs and peptide analogs utilizes a conventionally known method for introducing a dihydropyridine pyridinium salt-type redox moiety or a precursor thereof, for example, as described in International Publication No.
The method can be carried out by combining the methods described in 083/03968 and W085 / 03937 with well-known peptide synthesis methods. Finally, the quaternary forms of the amino acids and peptides are reduced to form the corresponding dihydropyridine compounds, which are used for administration, according to the methods described in the aforementioned Bodor U.S. Patent and the above-mentioned International Publication.

In a preferred embodiment of the present invention, a redox carrier system is selected as the redox system used in combination with the hydroxypropyl, hydroxyethyl, glucosyl, maltosyl or maltotriosyl derivative of β- or γ-cyclodextrin according to the present invention. I do. The drug portion and the carrier portion of the redox carrier system will be described in more detail later, but of course are described in detail in the above-mentioned various carrier patents and patent applications.
The selection of the appropriate drug and carrier moieties is not limited to the specific drugs and carriers disclosed in the above patents and patent applications and the present application, but the selected drugs and carriers are described in the above references. Any one can be selected as long as it meets the general requirements of the drug / carrier system as described.

As used herein, "drug" means any substance for use in diagnosing, curing, alleviating, treating or preventing a disease in a human or animal, or for enhancing a desired physical or mental development and condition. .

As used herein, a "centre acting" or "centrally acting" drug species, active ingredient or compound is defined as having a pronounced (usually major) pharmacological effect in the central nervous system (CNS) and in the brain. Needless to say, it means a kind of drug that occurs as a direct action of the drug.

Examples of such centrally acting drug classes include CNS
Systemic amines and other nervous system drugs (both sympathetic and parasympathetic) such as phenylethylamine (stimulant), dopamine (eg neurotransmitters used in the treatment of Parkinson's syndrome and hyperprolactinemia) And dopamine agonists), tyramine (stimulant), L-
Dopa (e.g., a dopamine precursor used to treat Parkinson's syndrome); muscle relaxants, tranquilizers and anti-depressants, e.g. Stabilizers, etc .;
Mild and powerful analgesics and narcotics; sedatives and hypnotics; anesthetic antagonists; vascular agents; stimulants; anesthetics; di-, tri-, tetra- and pentapeptides and other 2- Small peptides, such as small peptides containing 20 amino acid units, such as enkephalins (eg, Try-Gly-Gly-Phe-Leu) (enkephalins are not only analgesics, but also have an analgesic dose). Produces an antiepileptic effect in the brain at about 1/10 the dose); growth-promoting substances; antiepileptic and anticonvulsant drugs which generally include hydantoins such as phenytoin and ethotoin, and barbiturates such as phenobarbital; , Estradiol, testosterone, 17α
-Hormones, including steroid hormones such as ethynyltestosterone (ethisterone) (recent studies on hormonal sensitivity and histological mapping of specific steroid-binding cells in the brain have shown that steroids in the brain affect sexual behavior. The importance of action has been emphasized);
Amphetamine-like drugs; anticancer and antiparkinsonian drugs; antihypertensive drugs; -Tetrahydroacridine;
Antibacterials; centrally acting antihypertensives; centrally acting prostaglandins, eg PGD ₂ ; diagnostics, eg radiopharmaceuticals; monoamine oxidase (MEO), inhibitors; CN
S or brain important / essential amino acids such as tryptophan, which is both a nutrient and an antidepressant; as well as these analogous centrally acting compounds. For the purposes of the present invention, dopa and L-
Dopa (L-DOPA) is not classified as an amino acid, for example,
Classified as a CNS amine and dopamine agonist used in the treatment of Parkinson's syndrome.

Illustrative examples of other centrally acting drug classes are as follows: amphetamine, dextroamphetamine, levanfetamine, aletamine, cypenamine, fencamphamin, phenozolone, dilofuramine, methamphetamine, phenmetrazine, and phentermine ( These are sympathomimetic amines / cerebral stimulants and appetite suppressants); etriptamine (cerebral stimulants); codeine;
Oxycodone, pentazocine, anileridine, hydromorphone, morphine, and oxymorphone (these are narcotic analgesics); desipramine, nortriptyline, octtriptyline, maprotiline, opipramol, and protriptyline, which are used, for example, for endogenous depression Dibenzoazepine cerebral stimulants / tricyclic antidepressants; clonidine and methyldopa (these are sympathomimetics used, for example, for hypertension); piperidene, cyclimine, and procyclidine (these are centrally acting drugs). Anticholinergics; tranylcypromine (sympathetic cerebral stimulant / MAO inhibitor and antidepressant); acetophenazine, carfenazine, fluphenazine, perphenazine, and piperacetadine (these are phenothiazine tranquilizers) Benzoctamine (a sedative / muscle relaxant structurally similar to a phenothiazine tranquilizer); chlordiazepoxide, chlorazepate, nitrazepam, and temazepam (these are benzodiazepine tranquilizers); normacimethadol (methadone analgesic) Piminodin (a meperidine narcotic analgesic); tracazolate (a sedative / hypertensive drug); pridizirole (a centrally acting antihypertensive drug); sulpiride (an antidepressant / psychotropic drug); haloperidol and clopentixol Is a tranquilizer); norepinephrine (sympathomimetics / adrenergic agents); nalorphine and naloxone (antagonists); hydralazine (hypertensives); etotoin, phenobarbital, and aminoglutethimide (anticonvulsants); epinephrine (Adrenaline Drugs); Etamiban (bone marrow stimulant); Bimegurido (barbiturate such antagonists); amino phenazine tetrazole (stimulant); Yopidoru, iodo Pila set, iodo U plates (o
Iodohippuric acid), iodamide, and iopanoic acid (these are radioactive diagnostic agents); ephedrine, pseudoephedrine, oxymetazoline, and phenylephrine (these are sympathomimetic amines and decongestants); estradiol, estrone, and es Triols (natural estrogens); amoxicillin, oxacillin, carbenicillin, benzylpenicillin, phenoxymethylpenicillin, methicillin, officillin, ticarcillin, bacampicillin, episilin, hetacillin, pivanpacillin, methoxymethyl ester of hetacillin, and ampicillin antibiotics Amobarbital (an analgesic); Trihexyphenidyl (a centrally acting anticholinergic); hydroxyzine (a tranquilizer);
Chlortetracycline, demeclocycline, minocycline, doxycycline, oxytetracycline, tetracycline and metacycline (these are tetracycline antibiotics); flurazepam, bromazepam, dekimosepam, and lorazepam (these are benzocyazepine tranquilizers); Glutethimide (mild hypnotic / sedative); clindamycin, linocomycin, nalidixic acid, oxophosphoric acid, and phenazopyridine (antibacterial / antibiotic); betanidine and guanethidine (hypertensive / sympathetic blocker) Captopril (an antihypertensive drug); metiprilone (a mild hypnotic); amedalin, bupropion, cartazolate, daledarin, difluanine, fluoxetine, and nisoxetine (above the cerebrum)奮薬); propranolol (beta-blockers, antihypertensives); cloxacillin and dicloxacillin (penicillin antibiotics); butalbital (barbiturate sedative); GABA, .gamma. vinyl G
ABA, and γ-acetylene GABA (a neurotransmitter that may be used for epilepsy); valproic acid and its metabolites (5-hydroxy-2-n-propylpentanoic acid, 4-hydroxy-2-n-propylpentanoic acid) , 3
-Hydroxy-2-n-propylpentanoic acid, etc. (used as an antispasmodic); Valpromide (valproic acid derivative, used as an antispasmodic); Apomorphine (a narcotic inhibitor used in the treatment of photosensitive epilepsy / Emetics); forcodine (narcotic antitussive); methotrexate, mitoxantrone, podophyloxine derivatives (etopsid, teniposide), doxorubicin, daunamycin, and cyclophosphamide (cancer / antitumor agent);
Methylphenidate (stimulant); thiopental (anesthetic); ethinylestradiol and mestranol (estrogens); meptazinol, cyclazosin,
Phenazosin, Profadol, Methopon, Drocord, and Mifadol (these are narcotic analgesics); Buprenorphine, Narbuphine, Butorphanol, Levallorphan, Naltrexone, Nalmefene, Arazosin, Oxylorphan, and Nalmexone (these are narcotic antagonists or antagonists) Drugs); norgestrene and norethindrone (progestins); cephalothin, cephalexin, cefazolin, cefoxitin,
Mexalactam, ceforanide, cefloxazine, and cefapirine (cephalosporins); atenolol, nadolol, timolol, and metoprolol (beta blockers / hypertensives); ACTH (corticotropin or corticotropin) LHRH (a neurotransmitter that stimulates the secretion of the pituitary hormones LH and FSH, which acts on ovulation induction and fertility control / contraception); sulfadiazine and other sulfonamide antibiotics; ribavirin and Acyclovir (antiviral); chlorambutyl and melphalan (nitrogen mustard anticancer / antitumor drugs); methotrexate and aminopterin (these are antifolate anticancer / antitumor drugs); platinum Coordination complex, i.e., cis Datinomacin and mitomycin C (used for cancer chemotherapy); Thioguanine (purine / pyrimidine antagonist, used for cancer treatment); Vincristine and vinblastine (anti-cancer / anti-neoplastic drugs) Carcinogenic alkaloids); hydroxyurea and DON (anticancer urea derivatives); FSH, HCG, and HCS (pituitary and nonpituitary gonadotropins) (gonadotropins), for example, used in certain genital disorders N, N'-bis (dichloroacetyl) -1,8-octamethylenediamine (fertilisin) (agent for inhibiting male reproduction); lebruphanol (narcotic analgesic); benzestrol and diethylstilbestrol (synthesis) Estrogen); ethyl β-carboline-3-carboxylate (benzodiazepine antagonist); furosemide (diuresis / antihypertension) Hypertension drugs); dipyridamole and nifedipine (coronary vasodilator); and Purogabaido (GABA agonists and GABA precursor drugs).

Still other examples of centrally acting agents include non-steroidal anti-inflammatory / non-narcotic analgesics, such as propionic, acetic, fenamic, and biphenylcarboxylic acid derivatives. Specific examples of non-steroidal anti-inflammatory / non-narcotic analgesics that can be used in the present invention include ibuprofen, naproxen,
Flurbiprofen, Zomepirac, Sulundac, Indomethacin, Fenbufen, Fenoprofen, Indoproxen, Ketoprofen, Fluprofen, Bucloxic Acid, Tolmetin, Alclofenac, Fenglogic Acid, Ibufenac, Flufenisal, Pirprofen, Flufenam Acids, mefenamic acid, clonixeryl, clonixin, meclofenamic acid, flunixin, diclofenac, carprofen, etodolac, fendsal, prodric acid, sermethasin, indoxol, tetridamine, diflunisal, naproxol, piroxicam, methazamide, fluthiazine, and tesicam.

Preferred types of centrally acting drugs used in combination with redox carriers include central neurotransmitters, steroids, anticancer / antitumor drugs, antivirals, tranquilizers, memory enhancers, antihypertensives, sedatives, Antipsychotics and cerebral stimulants (especially tricyclic antidepressants).

Neurotransmitters include the amino acids mentioned above, such as GABA, GABA derivatives and other ω-amino acids, and glycine, glutamic acid, tyrosine, aspartic acid and other natural amino acids; dopamine, norepinephrine, and epinephrine. Catecholamines; serotonin, histamine and tryptamine; and neurotensin, luteinizing hormone-releasing hormone (LHRH), softstatin, enkephalins (met ⁵
- enkephalin and leu ⁵ - enkephalin, etc.), endorphins (.gamma., alpha-and β- endorphin), oxytocin M, and it can be exemplified Hepuchido such as vasopressin. Synthetic and semisynthetic analogs, such as analogs of LHRH in which one or more amino acids of LHRH have been eliminated and / or substituted with one or more other amino acids, which may be agonists or antagonists Included. For example, primary and secondary amine type LHRH
Similar substances are disclosed in U.S. Pat.Nos. 4,377,574; 3,917,825; 4,032,082
And 4,338,305.

Examples of steroids include hydrocortisone, betamethasone, cortisone, dexamethasone, flumethasone, fluprednisolone, meprednisone, methylprednisolone, prednisolone, prednisone, triamcinolone, cortodoxone, fludrocortisone, flulandrenolone acetonide (flurandrenolide), and paramethasone. Anti-inflammatory corticosteroids; testosterone and related analogs thereof, such as androgens such as methyltestosterone (17-methyltestosterone); and female hormones, including both estrogens and progestins, such as norgestrel , Norethindrone, norethynodrel, ethisterone, dimethisterone, allylestrenol, syngestol, ethine Down, progestins of lynestrenol, Norugesuteron, Norubinisuteron, ethynodiol, Okisogesuton, and soup, such as stalls,
And ethinyl estradiol, mestranol,
Estrogens such as estradiol, estriol, estrone, and kinesterol can be mentioned.

Ara-AC, pentostatin (2'-deoxycoformycin), Ara-C (silatabine), 3-deazaguanine, dihydro-5-azacitidine, thiazofurin, sangivamycin, Ara -A
(Vitarabin), 6-MMPR, PCNU, FENU, HENU and other nitrosoureas, spiromustine, bisbenzimidazole, L-alanosine (6-diaza-5-oxo-L-norleucine), DON, L-ICRF, TrimethylTM
M, 5-methyltetrahydrohomofolate, glyoxylic acid, sulfonylhydrazone, DACH, SR-2555, SR-250
8, desmethylmisonidazole, mitoxantrone, menogallol, aracinomycin A, filantside,
Bactobolin, affidocholine, homoharringtonine,
Levonantradol, acibicin, streptozotocin, hydroxyurea, chlorambutyl, cyclophosphamide, uracil mustard, melphalan, 5-FU (5
-Fluorouracil), 5-FUDR (Floxuridine),
Vincristine, vinblastine, cytosine arabinoside, 6-mercaptopurine, thioguanine, 5-azacytidine, methotrexate, adriamycin (doxorubicin), daunomycin (daunorubicin), largomycin polypeptide, aminopterin, dactinomycin, mitomycin C, and pododine Philotoxin derivatives such as etoposide (VP-16) and teniposide can be mentioned.

As antivirals, ribavirin; acyclovir (ACV); amantadine (may also be effective as an antiparkinsonian); 5-amidino-2- (5-amidino-2)
Diarylamidines such as -benzofuranyl) indole and 4 ', 6-diimidazolino-2-phenylbenzo (b) thiophene; 2-guanidino-4,5-di-n-propyloxazole and 2-guanidino-4,5-diphenyl 2-aminooxazoles such as oxazole; 6
[[(Hydroxyimino) phenyl] methyl] -1-
Benzimidazole analogs such as the syn and anti isomers of [(1-methylethyl) sulfonyl] -1H-benzimidazol-2-amine; 5,7-dimethyl-2-β
-D-ribofuranosyl-s-triazole (1,5-a)
Bridgehead C nucleosides such as pyrimidine; 2-deoxy-D-glucose, glucosamine, 2-deoxy-2-
Glucosides such as fluoro-D-mannose and 6-amino-6-deoxy-D-glucose; phenyl-
Phenylglucoside derivatives such as 6-chloro-6-deoxy-β-D-glucopyranoside; (S) -9- (2,3
-Dihydroxypropyl) adenine; thiozofurin; selenazofurin; 3-deazauridine; 3-deazaguanosine; DHPG; 6-azauridine; idoxlysine (5-iodo-2'-deoxyuridine); Zidovudine (AZT); dideoxycytidine;
And 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole.

As anticancer / antitumor and antiviral drugs, nucleoside derivatives (that is, purine or pyrimidine base structures having a substituent having one or more hydroxyl groups) are particularly useful. Compounds in this group include Ara-AC, pentostatin, Ara-C, dihydro-5-azacytidine, thiazofurin, sangivamycin, Ara-A, 6-MMPR, desmethylmisonidazole, 5-FUDR, cytosine arabic Noside, 5-azacytidine, ribavirin; acyclovir, (S) -9- (2,3
-Dihydroxypropyl) adenine, 6-azauridine, 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole, 5,7-dimethyl-2-β-D-ribofuranosyl-s-triazole (1,5- a) pyrimidine,
Compounds such as zidovudine (AZT), dideoxycytidine, dideoxyadenosine, dideoxyisosine, and DPHG.

Tranquilizers such as diazepam, oxazepam, lorazepam, chlordizepoxide,
Benzodiazepine tranquilizers such as flurazepam, bromazepam, chlorazepate, nitrazepam and temazepam; hydantoin tranquilizers / anticonvulsants such as phenytoin, etotoin, mephenytoin; acetophenazine, carphenazine, fluphenazine, perphenazine and piperanacetazine Tranquilizer; and the like.

Antihypertensives may include clonidine, methyldopa, betanidine, debrisoquine, hydralazine and guanethidine and analogs thereof.

Sedative, tranquilizing and antipsychotics can include many specific compounds of this type listed above, especially those of the phenocyanazine and benzodiazepine series, and analogs thereof.

Cerebral stimulants can include a number of the specific compounds listed above, especially sympathomimetic amine-type cerebral stimulants and tricyclic antidepressants, particularly preferred tricyclic compounds are dibenzoazepines and It is a similar compound.

Another example of a centrally acting drug class for use in combination with a redox carrier is a centrally active metabolite of a centrally acting drug. Representative examples of such metabolites include hydroxylated metabolites of tricyclic antidepressants, such as 10-hydroxynortriptyline, 2-hydroxyimipramine, 2-hydroxyimipramine,
E- and Z-isomers of hydroxydesipramine and 8-hydroxychloripramine; hydroxylated metabolites of phenothiazine tranquilizers such as 7-hydroxychlorpromazine; and desmethyl metabolites of N-methylbenzodiazepine tranquilizers such as Desmethyldiazepam. Other CNS useful in the present invention
Active metabolites will also be apparent to those skilled in the art. For example,
SL, an active metabolite of the GABA agonist progabide
-75102, and hydroxy CCNU, an active metabolite of CCNU, a nitrosourea anticancer drug. In general, these CNS active metabolites have been described as exhibiting activity in the scientific literature, but have never been administered as a drug per se. In many cases, the CNS activity of such active metabolites is determined by the drug (p
arent drug). However, these metabolites have not hitherto been administered as is because they cannot themselves cross the blood-brain barrier.

As described above, diagnostic agents including radiopharmaceuticals are also included in "agents acting centrally". Any diagnostic agent that can be derivatized to form a redox carrier system that passes through the BBB in reduced form and concentrates in the brain in its quaternary salt form and can be detected in the brain Is included. The diagnostic agent is "cold", ie, can be detected by X-rays (eg, a radiopaque agent) or by other non-invasive techniques such as mass spectrophotometry, NMR (eg, where the compound is C13, N15 , O18, S33, and S34
And the like when a stable isotope is contained). The diagnostic is also "hot", ie, radioactive iodine (I 123, I
It may be labeled with a radioisotope such as 125, I 131) and detected / imaged by radiation detecting / imaging means. Representative "cold" diagnostics that can form derivatives according to the present invention include o-
Examples include iodohippuric acid, iothalamic acid, iopidol, iodamide and iopanoic acid. Representative radiolabeled diagnostic agents include geohippuric acid (I 125, I 131), diotyrosine (I 125, I 131), o-iodohippuric acid (I 13
1), iothalamate (I 125, I 131), thyroxine (I 12
5, I 131), iotyrosine (I 131), and iodometaraminol (I 123) represented by the following structural formula.

In the case of a diagnostic agent, unlike a drug for treating a disease, a quaternary salt form "bound" in the brain is detected by video or other means, not the original original diagnostic agent itself. It becomes the form which is done. Furthermore, even for centrally acting drugs for the treatment or prevention of medical diseases,
For example, as long as it can be labeled with a radioisotope such as iodine or a stable isotope, such a label can be used after being converted into a diagnostic agent for incorporation into a dox carrier system.

As is evident from the known structure of many of the drug classes listed above, the drug of choice often has more than one reactive functional group, especially in addition to the group that binds the redox carrier. The drug may further contain other functional groups, such as hydroxyl, kilboxyl, amino, etc., which may be advantageously protected during synthesis and / or administration, if necessary.
Details of these protections are described in detail in the various patents and patent applications listed above. Such protected drug classes are, of course, also included in the definition of the term "drug" as used in the present invention.

As used herein, “dihydropyridine-type carrier”
Or [DHC], including, containing, or having a dihydropyridine nucleus, whether or not part of a larger basic nucleus, whether substituted or unsubstituted It will also be appreciated that any non-toxic carrier portion is meant. The only criterion is that it has the ability to cross the BBB and can be converted by oxidation in vivo to the corresponding quaternary pyridinium salt-type carrier [QC] ⁺ . As described above, the ionic pyridinium salt-type drug / carrier precursor [D-QC] ⁺ generated by this in vivo oxidation is prevented from flowing out of the brain, but is promoted from the general circulation. Is done. Thereafter, the covalent or equivalent bond linking the drug species [D] to the quaternary carrier [QC] ⁺ is metabolically cleaved, resulting in sustained release of drug [D] in the brain and the carrier moiety [QC] ] ⁺ Easy discharge occurs. A "covalent or equivalent bond" between such an agent and a quaternary carrier may be a simple direct chemical bond, such as, for example, an amide, ester, or any other similar bond, or even a thiazolidine bridge or peptide bond. It may be composed of a bonding group or a functional group. The latter linking group is generally required when the drug species used does not directly form a chemical bond with either the dihydropyridine or quaternary carrier. However, [D-QC] ⁺
And the bond [D-DHC] is meant to include, and is defined to include, all such substitutent bonds and linking groups. And simultaneously with the connected release of drug species [D] in the brain, the carrier part [QC]
Cleavage of the [D-QC] ⁺ type precursor, which results in easy excretion of ⁺ , occurs by specific enzymatic cleavage such as, for example, an esterase, amidase, cholinesterase, hydrolase, or peptidase.

As used herein, "a pharmaceutically acceptable non-toxic salt" refers to dihydropyridine, a reduced form of the redox carrier system or redox-like system formed by a pharmaceutically acceptable non-toxic inorganic or organic acid HX. It is meant to generally include non-toxic salts of the compounds. For example, the salt may be a salt derived from an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid;
Propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, fumaric acid And salts obtained from organic acids such as methanesulfonic acid and toluenesulfonic acid. In the present specification, for example, the `` anion of a non-toxic pharmaceutically acceptable acid '' used in connection with a pyridinium salt compound which is an oxidized form of a redox carrier system or a redox-like system refers to an inorganic or organic substance as described above. It is meant to include the anion of acid HX.

In the following description, the description “at least one reactive functional group selected from the group consisting of amino, hydroxyl, mercapto, carboxyl, amide, and imide” or a part of this description is used. The meaning of each functional group mentioned in this description is as follows.

"Amino" primary or secondary amino group, i.e. means a -NH ₂ or -NHR. Secondary amine groups are also referred to herein as -NH-. This is because the specific structure (strict type) of the R portion of the -NHR group is not particularly important, and R is a part of the drug residue D itself, and the drug is converted to a redox carrier system. This is because they remain unchanged.

 "Hydroxyl" refers to the group -OH.

 “Carboxyl” means a —COOH group.

 "Mercapto" refers to the -SH group.

The term "amido", carbamoyl (-CONH ₂₎ or substituted carbamoyl (-CONHR) or a sulfamoyl (-SO ₂ NH ₂₎ or substituted sulfamoyl (-SO ₂ NHR)
Means a functional group. -CONHR group and -SO ₂ NHR group may be referred respectively -CONH- and -SO ₂ NH- and herein. This is because the specific structure of the R moiety is not important, as R is part of the drug residue D itself and remains unchanged when the drug is converted to a redox carrier system.

"Imido" means a functional group represented by the following structure, This structure is a structure characterizing an imide compound (that is, a compound having a succinimide type or phthalimide type structure).

A number of different dihydropyridine pyridinium salt type redox carrier moieties are disclosed in the aforementioned carrier patents or published patent applications. The following is a list of representative major classes of dihydro compounds and the corresponding quaternary salts, which are exemplary and
It is not intended to be exhaustive.

(1) When the drug has at least one hydroxyl or mercapto or primary or secondary amino functional group: by replacing a hydrogen atom in at least one of the functional groups with any of the following [DHC] groups: To combine.

In the above formula, the dotted lines in formulas (a ′), (b ′) and (c ′) indicate that a double bond exists at either the 4-position or the 5-position of the dihydropyridine ring; (d ′) ,
(E ') and (f' dotted in) indicates that the double bond in either the 2-position or 3-position of the dihydroquinoline ring is present; R ₁ is C ₁ -C ₇ alkyl, C ₁ ~ C ₇ haloalkyl or C ₇ -C ₁₀ aralkyl group; R ₃ is C ₁ -C ₃
Means an alkylene group; X is -CONR'R "(R 'and R" may be the same or different, each means H or C ₁ -C ₇ alkyl group) or a or X, is -CH = be NOR (R means H or C ₁ -C ₇ alkyl group); X substituent in formula (a ') and (c') a carbonyl-containing group and the formula in (b '), respectively the dihydropyridine (D ′) and (f ′) can be attached at the 2-, 3- or 4-position of the ring;
And the X substituent in the formula (e ') are, respectively, 2-position, 3-position of the dihydroquinoline ring.
And the carbonyl-containing group in formulas (g ′) and (j ′) and the X substituent in formula (h ′) can be bonded to the 1-, 3-positions of the dihydroisoquinoline ring, respectively. Alternatively, it can be bonded to the 4-position.

(2) When the drug has at least one carboxyl functional group: The drug is bound by replacing a hydrogen atom in at least one of the carboxyl functional groups with any of the following [DHC] groups.

(A) When the compound has one or more —COOH groups available for derivative formation: In the above formula, the dotted lines in formulas (i ′), (ii ′) and (iii ′) indicate that a double bond exists at either the 4-position or the 5-position of the dihydropyridine ring, and the formula (i
v '), (v') and (vi 'dotted in) indicates that a double bond is present either 2-position or 3-position of the dihydroquinoline ring; Z' is C ₁ -C ₈ straight chain or branched chain alkylene group, preferably denotes a C ₁ -C ₃ straight or branched chain alkylene group; Q is -O- or -NH-; R
₁ is C ₁ -C ₇ alkyl, C ₁ -C ₇ haloalkyl or C ₇ -C
_{10 represents an} aralkyl group; R ₃ represents a C ₁ -C ₃ alkylene group; X represents —CONR′R ″ (R ′ and R ″ may be the same or different and are each H or C ₁ -C _7; Or X represents -CH = NOR (R
Be a means H or C ₁ -C ₇ alkyl group); the formula (i ') and (iii' X substituent in carbonyl-containing group and the formula in) (ii ') the 2-position of the respective said dihydropyridine ring Carbonyl-containing groups in formulas (iv ′) and (vi ′) and the X substituent in formula (v ′) can be attached to the 2-, 3-positions of the dihydroquinoline ring, respectively. Or at the 4-position; and formulas (vii ') and (i
x ') and a carbonyl-containing group of formula (viii')
X can be engaged at the 1-, 3- or 4-position of the dihydroisoquinoline ring, respectively.

(B) When there is only one -COOH group available for derivative formation: In the above formula, the dotted line in the formula (xii ') indicates that a double bond exists at either the 4-position or the 5-position of the dihydropyridine ring; the dotted line in the formula (xiii') indicates that the dihydroquinoline ring has Indicates the presence of a double bond at either position 2 or position 3; Means the skeleton of the sugar molecule; n ^iv means a positive integer equal to the total number of -OH groups in the sugar molecule used to derive the skeleton; n ^v denotes the sugar used to derive the skeleton. Means a positive integer one less than the total number of -OH groups in the molecule;
A in each of i '), (xiii') and (xiv ') independently represents a hydroxyl group or D', where D 'is a centrally acting group containing one reactive carboxyl function. (X ') and (x') means the residue of the drug of formula (I) wherein one hydrogen atom is removed from the carboxyl function of the drug.
i ′) each R ₄ is independently a hydroxyl group, Or D '[where the scores have the same meaning as described for structures (xii') and (xiii ');
D 'has the same meaning as described for structures (xii'), (xiii ') and (xiv'); R ₁ is C ₁ -C ₇ alkyl, C _1
1 -C ₇ means haloalkyl or C ₇ -C ₁₀ aralkyl group; displayed carbonyl group, the pyridinium or quinolinium 2-position of the ring, 3- or 4-position, or isoquinolinium 1-position of the ring, 3- or 4 Where the structures (x ′) and (xi ′)
At least one R _{4 in} each of (Where R ₁ , the dotted line, and the position of the carbonyl-containing group are as described above); and provided that two or more R ₄ groups present in the compound are the carbonyl-containing group described above. Is that all such carbonyl-containing groups in this compound are the same.

(3) When the drug has at least one -NH-functional group that is part of an amide or imide structure, or at least one primary or secondary amine functional group with a low pKa: at least one of the functional groups The drug is attached by substituting one of the hydrogen atoms with one of the following [DHC] groups.

In the above formula, R is hydrogen, C ₁ -C ₇ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₇ haloalkyl, furyl, phenyl group, or one or more halogen, lower alkyl, lower alkoxy, A phenyl group substituted with a carbamoyl, lower alkoxycarbonyl, lower alkanoyloxy, lower haloalkyl, mono (lower alkyl) carbamoyl, di (lower alkyl) carbamoyl, lower alkylthio, lower alkylsulfinyl or lower alkylsulfonyl group; The dotted lines in k '), (l') and (m ') represent the 4
Indicates the presence of a double bond in either position or position 5; the dashed line in formulas (n '), (o') and (p ') indicates that the dihydroquinoline ring is in either position 2 or position 3. Indicates that a double bond is present; R ₁ represents a C ₁ -C ₇ aralkyl, C ₁ -C ₇ haloalkyl or C ₇ -C ₁₀ aralkyl group; R ₃ represents a C ₁ -C ₃ alkylene group. Means; X is -CON
R'R "(R 'and R" may be the same or different, mean H or C ₁ -C ₇ alkyl groups, respectively)
Or X is -CH = NOR (R is H or C
₁ -C ₇ is an alkyl group means a); Formula (k ') and (m' X substituent in carbonyl-containing group and the formula (l ') in) the 2-position of the respective said dihydropyridine ring, 3- or The carbonyl-containing group in formulas (n ') and (p') and the X substituent in formula (o ') can be attached at the 2-, 3-, or 4-positions of the dihydroquinoline ring, respectively. And the carbonyl-containing group in formulas (q ') and (s') and the X substituent in formula (r') are each in the 1-, 3-, 3-position of the dihydroisoquinoline ring.
Or at position 4.

Agents containing a secondary or tertiary hydroxyl function can be attached to any of (k ') to (s') in the [DHC] group described above. in this case, The moiety is derived from an aldehyde RCH ₂ O (eg, chloral, acetaldehyde, formaldehyde, or benzaldehyde) that can react with the agent to form the corresponding hemiacetal.

The following is a list of dihydropyridine pyridinium salt forms that can be used as pharmaceuticals in parenteral pharmaceutical compositions in combination with certain cyclodextrin derivatives in accordance with the present invention, particularly for brain targeted drug delivery purposes. Redox carrier system (this may be also referred to as chemical supply system or CDS in this specification)
Is an example of a dihydropyridine compound which is a reduced form of

The cyclodextrins used in the present invention are the hydroxypropyl, hydroxyethyl, glycosyl, maltosyl and maltotriosyl derivatives of β-cyclodextrin, and the corresponding derivatives of γ-cyclodextrin. The hydroxyalkyl group can contain one or more hydroxyl groups such as, for example, hydroxypropyl, dihydroxypropyl, and the like.
Glucosyl, maltosyl and maltotriosyl derivatives can contain one or more sugar residues, for example, glucosyl or diglucosyl, maltosyl or dimaltosyl and the like. Various mixtures of cyclodextrin derivatives (eg, mixtures of maltosyl and dimaltosyl derivatives) can also be used. Specific examples of cyclodextrin derivatives useful in the present invention include:
Hydroxypropyl-β-cyclodextrin (HPCD or HPBCD), hydroxyethyl-β-cyclodextrin (HEBCD), hydroxypropyl-γ-cyclodextrin (HPGCD), hydroxyethyl-γ-cyclodextrin (HEGCD), dihydroxypropyl-β - cyclodextrin (2HPBCD), glucosyl -β- cyclodextrin (G ₁ -β-CD or G ₁ BCD), diglucosyl -β- cyclodextrin (2G ₁ -β-CD or 2G ₁ BC
D), maltosyl-β-cyclodextrin (G ₂ -β-C
D or G ₂ BCD), maltosyl-γ-cyclodextrin (G ₂ -γ-CD or G ₂ GCD), maltotriosyl-β-
Cyclodextrin (G ₃ -β-CD or G ₃ BCD), maltotriosyl -γ- cyclodextrin (G ₃ -γ-CD or G ₃ GCD), and dimaltosyl -β- cyclodextrin (2G ₂ -β-CD Or 2G ₂ BCD), and maltosyl-β-cyclodextrin / dimaltosyl-β-
Mixtures of the above, such as a mixture of cyclodextrins.

The hydroxypropyl-β-cyclodextrin used in the method of the present invention is commercially available. Alternatively, this can be carried out in a known manner, especially in the International Journal of Pithu et al.
of Pharmaceutics , 29 , 73-82 (1986). In the above document
A typical synthesis method using the method of Pitha et al. Is shown below.

31 g of sodium hydroxide was dissolved in 250 ml of water in the flask. Next, 100 g of β-cyclodextrin was added, and the solvent was heated to form a solution. The flask was cooled and 50 ml of propylene oxide was added. During this addition, the flask was fitted with a dry ice / acetone cooler. The resulting solution was warmed to room temperature and stirred for 72 hours. The solution was then neutralized with concentrated hydrochloric acid and diluted with water. The solvent was removed under reduced pressure, and the resulting syrupy residue was dissolved in ethanol. After stirring at room temperature for 30 minutes, the precipitated sodium chloride was removed by filtration. The filter cake was washed with ethanol, and the ethanol layers were combined and concentrated under reduced pressure. The residue was dissolved in water, and the cellulose acetate membrane (No. 7, 38 mm, 4.6 ml / cm, molecular weight separation = 1000, Fi
(Sher Scientific). After 5 hours at 0 ° C., the solution was removed from the dialysis tube and freeze-dried. The obtained solid was suspended in acetone and stirred overnight. The solid obtained by filtration was resuspended in acetone and stirred for 24 hours. The solid was filtered off, dissolved in 200 ml of water and lyophilized. 7
5 g of purified hydroxypropyl-β-cyclodextrin were obtained. The degree of substitution was calculated by comparing the NMR analysis results with authentic samples.

A 50% solution (w / w) of 2-hydroxypropyl-β-cyclodextrin (HPCD) was prepared using distilled water to form an inclusion compound with E ₂ -CDS (estradiol-CDS). . After addition of excess E ₂ -CDS, the solution was purged with helium. The resulting suspension was then sonicated for 30 minutes, filtered through a glass filter (ASTM 10-15M, Pyrex No. 36060) and lyophilized overnight. E ₂
Best results were obtained when the CDS / HPCD clathrate aqueous solution was strongly frozen for at least 10 hours and then lyophilized. A small amount of the dried clathrate was dissolved in methanol and analyzed by high pressure liquid chromatography (HPCD) to measure the degree of clathrate formation. The degree of formation of the clathrate thus determined was in the range of 20 to 40 mg / g, and the solubility of the clathrate obtained was 2.2 × 10 ⁴ mg / g.

The above Pith for preparing HPCD by condensing propylene oxide with β-cyclodextrin in an alkaline aqueous solution
The method of a et al. unfortunately has the disadvantage that it requires a number of steps, especially for the purification of the product. After completion of the condensation reaction, the reaction mixture was neutralized with hydrochloric acid, water was evaporated under reduced pressure, and the syrupy residue was dissolved in ethanol.
The main by-product of the reaction, sodium chloride, is precipitated.
After filtration, the ethanol is evaporated under reduced pressure, the residue is dissolved in water and dialysis is carried out to remove residual sodium chloride and propylene oxide polymerization products. During dialysis,
Some of the produced hydroxypropyl-β-cyclodextrin passes through the membrane and is lost. The dialysate is then lyophilized, stirred twice in acetone and washed to remove residual polymerization products. Finally, the hydroxypropyl-β-cyclodextrin is freeze-dried again. This second lyophilization is necessary because the product after washing with acetone is not homogeneous.

To address the above difficulties associated with the Pitha et al. Method, a new method for the synthesis of HPCD was developed by M. Smulkowski of the University of Florida. The new method involves removing sodium hydroxide from the reaction mixture with an ion exchange resin (H ⁺ form). As a result, many of the time-consuming purification steps of the Pitha et al. Method can be avoided. Further, the amount of sodium hydroxide used can be reduced from 7 equivalents per 1 equivalent of β-cyclodextrin used in the method of Pitha et al. To 2 equivalents of sodium hydroxide per 1 molecule of cyclodextrin, and it is possible to reduce the amount of sodium hydroxide. A product with NMR properties and optical rotation is formed.

According to the newly developed method of synthesizing HPCD, first, β-cyclodextrin is condensed with propylene oxide in an alkaline solution, and sodium hydroxide is ion-exchanged using an ion exchange column (Dowex 50W-X8, H ⁺ type). Remove
The eluate was evaporated under reduced pressure and concentrated to 1/2 of the original volume,
The remaining solution is lyophilized and the white solid obtained is washed with acetone, lyophilized again, crushed and sieved. Possible modifications of this method include (1) neutralization using an ion exchange resin in a reaction flask, washing the resin in a filter funnel after filtering off, and (2) dissolving the cyclodextrin. Use calcium hydroxide, magnesium, lithium or potassium. (3) Remove the hydroxide after the reaction by saturating the reaction mixture with carbon dioxide or neutralizing with sulfuric acid instead of an ion exchange resin. (4) further reducing the amount of sodium hydroxide used (1-2 equivalents), and (5) omitting the second freeze-drying.

Next, representative operations using the newly developed improved method will be described.

Dissolve 3.53 g of sodium hydroxide in 751 ml of water and add β-
50 g of cyclodextrin were dissolved and treated at 0 ° C. with 29 ml of propylene oxide. The reaction mixture was kept at this temperature for 5 hours and then at room temperature for 42 hours. After this time has elapsed, the reaction fit is applied to a Dowex 50W-X8 column (H
⁺ ), Wash the column with water, and elute 100 ml volume
After evaporating under reduced pressure until freezing, freeze-drying was performed. The obtained white solid was washed with acetone to obtain 51 g of HPCD,
It showed the same degree of substitution (4.7) and NMR spectrum as HPCD prepared by the method of Pitha et al. Ignition residue is 0.0
%Met. The optical rotation was also identical to the product of Pitha et al.

Similar results were obtained in a condensation experiment using 7.71 g of sodium hydroxide for 25 g of β-cyclodextrin.

In this new improved synthesis of HPCD, purification of the solution using activated carbon before the final freeze-drying further improves the results. That is, when the aqueous solution eluted from the ion exchange column of Dowex 50W-X8 is treated with activated carbon, HP
Since most of the polymerization product was removed without loss of CD, the filtrate, which was only washed once with ethyl acetate, could be finally freeze-dried as it was. This way,
Only one freeze-drying was needed. At least on a small scale, crystallization (crystallization) of the final product is also possible instead of freeze-drying.

This modified new method (using activated carbon)
The product obtained by the new method before modification and Pi
It appears to be superior to the product obtained by the method of tha et al. in the following respects. First, in the modified method, the product is pure white and forms a colorless aqueous solution, whereas the product of the previous two methods forms a yellow aqueous solution. Second, the product of the modified method is not oily. This may be due to the removal of more lightly substituted, less water soluble, oily cyclodextrins.

HPCD can be prepared with various degrees of substitution, such as 5 to 7. Typically, HPCD (AS
Generate DS 7). The mass spectrum of the obtained isomer mixture of HPCD has a substitution degree of 7 as a center. This spectrum "gentle" the sample using fast atom bombardment
Obtained by ionization. The spectra generated are similar to those already reported (obtained by californium 252 plasma desorption), both in terms of the symmetry of the isomer distribution and the numerical distribution of the isomers formed.

Hydroxyethyl-β-cyclodextrin (HEBC
D) can be prepared in a similar manner to HPCD using the improved synthesis method of HPCD detailed above, substituting an equivalent amount of ethylene oxide for the propylene oxide used therein.

2-Hydroxypropyl-γ-cyclodextrin (HPGCD) can also be synthesized in the same manner using the same basic operation as in the case of HPCD, by replacing β-cyclodextrin as a raw material with γ-cyclodextrin.
However, for 7 β-cyclodexolins,
Since cyclodextrin contains eight glucose residues, the amount of propylene oxide used can be reduced to reduce the degree of substitution. Using 0.75 mol of propylene oxide (about 20% excess considering 8 OH groups) per 0.077 mol of γ-cyclodextrin gives HPGCD with a degree of substitution of 8, and 0.56 mol of propylene oxide (about 10 % Less) gives a degree of substitution of about 7.

Hydroxyethyl-γ-cyclodextrin (HPGC
D) can be synthesized by simply using an equivalent amount of ethylene oxide instead of propylene oxide in the same manner as the above-mentioned method of synthesizing HPGCD.

Thus, the hydroxyalkylcyclodextrins used in the present invention can be prepared by the method of Pitha et al. Or a modification thereof. When synthesized in this manner, the resulting cyclodextrin derivative is essentially an amorphous mixture. The importance of the amorphous (amorphous) nature of cyclodextrins has been demonstrated by Pitha et al., JP
harm . Sci ., Vol. 74, No. 9, September 1985, 987-990. The advantage of the amorphous nature of this material becomes more pronounced as the concentration of cyclodextrin is higher.

 Another group of cyclodextrins used in the present invention,
That is, the glucose of β- and γ-cyclodextrin
, Maltosyl and maltotriosyl derivatives
Is highly water-soluble compared to cyclodextrin
Cyclodextrins. This branched cyclodext
Phosphorus is microbiologically derived from the starting material cyclodextrin.
Can be manufactured by a standard method. Glucosyl-β-
Cyclodextrin is available from Bacillus auvensis (Baci
llus ohbensis) cyclomaltodextrin glucano
Large scale β-cyclodextran by transferase
Can be obtained from the mother liquor of phosphorus synthesis; Koizumi et al.Che
m.Pharm.Bull.,35(8), 3413-3418 (1987) and
See the references cited in Martosil and Ma
Lutotriosyl-β- and γ-cyclodextrin
Is the raw material cyclodextrin and maltose or maltose
Pseudomonas from Totriose
Isoamylase or Klebsiella aerogenes
(Klebsiella aerogenes) by the reverse action of pullulanase
Glucosyl-γ-cyclodexto can be prepared
Phosphorus is an enzymatic addition of maltosyl-γ-cyclodextrin.
Can be prepared by hydrolysis; Okada et al.Chem.Pha
rm.Bull.,36(6), 2716-2158 (1988) and its contents
See the cited references. In the presence of pullulanase
Reacting lactose with β-cyclodextrin
The production of maltosyl-β-cyclodextrin by
Also described in JP-A-61-287902 and JP-A-61-197602.
It is listed. Maltosyl-β-cyclodextrin and
And various dimaltosyl-β-cyclodextrins
Mixtures, such as IS commercially available from Ensuiko Sugar Co., Ltd.
OELEAT Can be used without any problems.

With the development of a carrier-mediated dihydropyridine pyridinium salt redox system, which in its dihydropyridine form is also called a chemical delivery system (CDS),
Increased and / or sustained supply of various drugs to the central nervous system has become possible. Although the physicochemical properties of this CDS are optimal for absorption and retention in the brain, it is often unsuitable for aqueous formulations or use as aqueous solutions. Its remarkable example is E ₂ -CDS, i.e. CDS derived from estradiol. The dihydro nicotinate (E ₂ -CDS) passes through the BBB, it is oxidized to E ₂ Q ⁺ quaternary salt which corresponds in the brain. E ₂ Q ⁺ thus generated
Is sustained in the brain, from which estradiol is gradually released, which exerts a significant central estrogenic effect. These effects include LH (luteinizing hormone) in ovarectomized rats.
Suppression, and reversible suppression of the periodicity of such non-castrated female rats, these effects are long lasting. E ₂ -CDS is highly lipophilic and very low in water solubility (0.2 μg / ml). Therefore, it was necessary to dissolve E ₂ -CDS in a water-miscible organic solvent such as dimethylsulfoxide (DMSO) or dimethylacetamide (DMA) before administration. This procedure is not necessarily inappropriate in laboratory animal experiments, but is obviously inappropriate when administered to humans, as listed above. Therefore, development of an aqueous composition or aqueous solution of E ₂ -CDS has been studied. The request criteria for such compositions, it is less toxic, the effect of the supply to the E ₂ Q ⁺ in the brain is equivalent to that of the DMSO or DMA solution of E ₂ -CDS, and development techniques Can also be applied to other dihydropyridine pyridinium salt redox systems.

Next, details of experimental examples and experimental results performed for verifying the effects of the present invention will be described.

Experimental Example Materials Used 3-hydroxy-17β-[(1-methyl-1,4-dihydropyridin-3-yl) carbonyl] oxyestra-1,3,5 (10) -triene (E ₂ -CDS), 1- Methyl-3
-[N- {β- [3,4-bis (pivalyloxy) phenyl] ethyl} carbamoyl] -1,4-dihydropyridine (DA-CDS), 17β-[(1,4-dihydro-1-methyl-
3-pyridinylcarbonyl) oxy] androst-4
- en-3-one (T-CDS _1), 1- methyl--3-N-
[3- (benzyloxycarbonyl) propyl] carbamoyl-1,4-dihydropyridine (GABA-CDS ₁ ), 1-
Methyl-3-{[2- (9-guanylmethoxy) ethoxy] carbonyl} -1,4-dihydropyridine (ACV-CD
S), and 17β-{[(1-methyl-1,4-dihydropyridin-3-yl) carbonyl] oxy} -19-norpregn-4-en-20-yn-3-one (N-CDS)
Was synthesized by a known method.

2-Hydroxypropyl-β-cyclodextrin (HPCD) (statistical average degree of substitution = 5.1 or 7) was prepared by the method of Pitha et al. Other cyclodextrins (α,
β or γ) is obtained from Aldrich Chemical Co.
Other steroids (estradiol, estradiol 17-valerate, estriol, estrone, estradiol 3-methyl ether and testosterone 17-propionate) were purchased from Sigma Chemical Co.

All prepared compounds were well confirmed by spectroscopy and microcombustion analysis (Atlantic Microlabs) prior to the experiment. Mass spectroscopy was performed using a Kratos MS80RFA bifocal instrument equipped with a fast atom gun. The cyclodextrin mixture was analyzed by fast atom bombardment of a sample prepared in a glycerol matrix. The degree of substitution was determined from the isomer mass distribution. Nuclear magnetic resonance spectra (NMR) were obtained on a Varian EM360 60 MHz spectrometer. The measured value is the internal standard [3- (trimethylsilyl)
Propionic 2, 2, 3, 3-d4 acid, sodium salt, was recorded for DDS], all samples were measured in D ₂ O. The degree of substitution was calculated by comparing the integrated area attributed to anomeric hydrogen with that attributed to the hydroxypropyl functional group.

Effect of dissolving agent Excess E_Two-CDS for 30 minutes with an aqueous solution of a suitable dissolving agent
Sonicated. Then, the obtained suspension is centrifuged.
Made of polyvinylidene difluoride (PVDF) with a pore diameter of 0.45μm
Filtration membrane [Millex-HV4, Millipore ], And HP
Analysis was performed by LC (high pressure liquid chromatography). 2
-Hydroxypropyl-β-cyclodextrin (HPC
In experiments using D), excess E_Two-CDS at various concentrations (% w /
v) was added to the HPCD, and the solubility (mg / ml) was analyzed by spectrophotometry (UV =
360 nm, ε = 6487 in methanol). The entire
The approximate value of the equilibrium constant for_Two-Mmol concentration of CDS
The relationship between the degree and the millimolar concentration of added cyclodextrin
Asked by the staff. Millimolar concentration of cyclodextrin
Is the average fraction of the mixture of isomers determined by mass spectrometry
It was calculated using the child size. In addition, a 50% w / w solution of HPCD
The dissolution effect is determined by a series of steroid and dihydropyridine
The combination (CDS) was examined. These experiments are also described above.
Was performed in the same manner as described above.

Preparation excess E ₂ -CDS or other CDS solid inclusion compound was added to 50% w / w solution of HPCD. The resulting suspension was sonicated for 30 minutes, filtered through a 0.45 μm PVDF filtration membrane, and lyophilized. The degree of contamination (inclusion degree) was measured by spectrophotometry or HPLC. In some cases, the effect of the lysing agent on the degree of contamination was investigated. This is because a small amount of polyoxyethylene 20 cetyl ether (Br
ij), polyoxyethylene sorbitan monooleate (Tween 80) or ethanol was added.

Analytical Methods For spectrophotometric concentration determination, Cary 219 (Varia
n) or HP 8415A Diode Array (Hewlett Packard)
A spectrophotometer was used. A standard curve in methanol was generated, which gave a correlation coefficient greater than 0.999. The monitoring wavelength was 360 nm for CDS, and 220 nm was used for estrogens.

The HPCD device is an Autoch with an Rheodyne injector
rom M500 pump or Perkin-Elmer series 4 pump, Kratos Spectroflow 757 variable wavelength detector, and Beckman recorder or LCI-100 integrator (P
erkin-Elmer). Separation, An
The analysis was performed on an analytical column of alytical Sciences, Inc. (ASI), particle size 10 μm, C18 reverse phase, 30 cm × 3.9 mm (inner diameter). The flow rate was 16 ml / min, the compound was detected at 360 nm, and the temperature was room temperature in all measurements. 82: 1: 1: 16 acetonitrile: tetrahydrofuran: acetic acid: mobile phase containing water, at 4.4 minutes E ₂ -CDS at 4.4 min DA-CDS, the T-CDS ₁
At 6.8 minutes, the N-CDS was eluted at 5.2 minutes each. GABA
For -CDS ₁ , a mobile phase consisting of a 50: 1: 1: 48 mixture of the same components was required. Retention time was 5.2 minutes. Other compounds were analyzed spectrophotometrically.

It restrained Sprague-Dawley rats (female, body weight 200g) with animal experiments awareness, DMSO solution state E ₂ -CDS15mg / kg or inclusion compounds of _{HPCD, (E 2 -CDS-HPCD} ) form an aqueous solution of of E ₂ -CDS 5 mg / kg, it was administered by intravenous injection (tail vein). At various times after dosing, animals were dissected and vascular blood and organs were collected. Then weigh each organ,
Homogenized in water and deproteinized with cold acetonitrile. The obtained organ homogenate is centrifuged, and the supernatant is subjected to a precolumn concentration method (precol
E ₂ Q ⁺ and E ₂ − using the umn enrichment technique).
Analyzed for CDS.

Results and Discussion As mentioned above, cyclodextrins have been used to enhance aqueous solutions of a number of drugs, including steroids. The cyclic oligomer has different numbers of α-1.4-
Contains bound glucose units. The number of these units (α = 6, β = 7, γ = 8) determines the size of the conical cavity (internal space) that can be used to contain a large number of drugs. The stability of the formed inclusion compound depends on the degree of fitting of the drug into the cyclodextrin (fitting property) and the concentration of the cyclodextrin. Unfortunately, β-cyclodextrin, the most suitable cyclodextrin for inclusion compound formation with steroids, has low water solubility. This property is derived from the high degree of hydrogen bonding that occurs within the crystal lattice. In addition to this problem, β-cyclodextrin is known to cause renal damage in rats. This toxicity is also due, at least in part, to poor water solubility. In any case, the water solubility of E ₂ -CDS is
In the case of equilibrium in a solution having various concentrations of-, β- or γ-cyclodextrin, little change was observed. As shown in Table 1 below, 50
The α- cyclodextrin concentrations up mM does not increase only 25 times of the water-soluble E ₂ -CDS, only increased to 135-fold and 110-fold respectively water of this CDS in the β- and γ- cyclodextrin Met. The relationship between the water solubility of E ₂ -CDS and the concentration of unsubstituted cyclodextrin was not linear, which was the same situation as observed when the drug substance, steroid, was used as an inclusion compound. In any case, for α-, β- or γ-cyclodextrin,
The resulting low water solubility and relatively low inclusion compound formation are unsuitable for drug development purposes. The toxicity of β-cyclodextrin further strengthens this assessment.

Some attempts have been made to increase the water solubility, and thus the utility, of cyclodextrins. Although various methylated derivatives have been reported, the acute toxicity of this modified compound is generally greater than the parent compound (unsubstituted cyclodextrin). More recently, it has been reported that hydroxypropylation of β-cyclodextrin yields amorphous cyclodextrin derivatives. The product, 2-hydroxypropyl-β-cyclodextrin (HPCD), is a mixture of isomers that can be described by the average degree of statistical substitution (ASDS). This ASDS value can be determined using NMR or mass spectrometry. This highly water-soluble mixture of isomers has been shown by Pitha et al. To significantly increase the solubility of a number of compounds, including gonadal steroids. In addition, preliminary toxicity studies have shown that side effects, if any, after oral or intravenous administration are minimal.

HPCD (ASDS 5.1 or 7) was prepared according to the method of Pitha et al. The mass spectrum of the isomer mixture of HPCD was centered around a substitution degree of 7. This spectrum was obtained by "mildly" ionizing the sample using fast atom bombardment. The spectra obtained were similar to the previously reported spectra (obtained from Californium-252 plasma desorption), both in the symmetry of the isomer distribution and in the numerical spread of the isomers formed. In the cited example, no toxic unsubstituted β-cyclodextrin was detected, even when the degree of substitution of ASDS 5.1 was low.

When applying this HPCD to E ₂ -CDS, HPCD with a low ASDS was selected. As the degree of substitution increases, the tendency of cyclodextrins to form inclusions, possibly due to steric interactions, decreases, and the surface activity of the resulting inclusions increases. In general, increasing surface activity is undesirable because increased surface activity also increases the propensity of the substance to cause hemolysis. ASDS 5.1 and 7
Any of HPCD, and showed a marked effect on the solubility of E ₂ -CDS. In the case of ASDS 7, E ₂ with increasing concentration of HPCD
-A linear increase in the solubility of CDS (r = 0.959) was clearly shown. 30.2 mg / ml could be dissolved at an HPCD concentration of 62.5% w / w. ASDS 5.1 HPCD with 62.5% w / w concentration
It was possible to dissolve the E ₂ -CDS of 35 mg / ml. HPCD with lower ASDS gave a 15% increase in contamination. These data, 5-digit ones compared to the solubility in water of E ₂ -CDS (1
This reflects the result of an increase in solubility (by a factor of 50,000) (see Table 1). Plotting the data obtained from the ASDS 7 HPCD experiment as the mmol concentration of added ECD (calculated based on the average molecular weight of the mixture of isomers) versus the mmol concentration of dissolved E ₂ -CDS gives a straight line with a slope of 0.2. Became. This slope serves as a criterion for evaluating the overall stability of the cyclodextrin inclusion compound, and serves as a suitable material for comparison with other systems.

The resulting solution was lyophilized to a solid inclusion compound. 50% w / w solution of HPCD is uniquely E ₂ -CDS of 37mg for inclusion compound 1g solid (contaminated degree 37mg / g)
Occurred. This clathrate was stable as a dry powder and could be easily returned to solution with water. In these operations, it was important to maintain the concentration of the HPCD component at 20% w / v or more. If the concentration is lower than this, precipitation may occur. Various additives (dissolving agents) such as Brij (0.7
% W / w), Tween 80 (0.8% w / w) or ethanol (10%
% V / v), several attempts were made to increase the degree of inclusion of the entrapment compound. 189m contamination by adding Brij
g / g, but the obtained clathrate was not stable, and the contamination decreased to 42 mg / g in 12 days. The remaining solubility was not very effective. Therefore, the upper limit of the degree of contamination for the formation of a stable clathrate is about 40 mg in the above example.
/ g.

Since the inclusion compound is formed between the E ₂ -CDS and the various isomers component cyclodextrin mixture, E ₂ -CD
Some of S rapidly without dissociation is likely to reduce the biologically effective concentration of E ₂ -CDS. To investigate this possibility, HPCD (ASDS 5.1) in pharmaceutical compositions containing _{E 2 -CDS (E 2 -CDS-} HPCD) measures the ability to provide E ₂ Q ⁺ to the brain, E _2- CDS dissolved in DMSO and administered
Compared to ₂ Q ⁺ supply. One of E ₂ -CDS of 5 mg / kg of E ₂ -CDS or HPCD aqueous solution of 15 mg / kg in DMSO after systemic administration were determined brain levels of E ₂ Q ^+. Consider the difference in dose, i.e., the case shown data as dose% per g, and after the administration of E ₂ -CDS-HPCD at E ₂ after administration of -CDS and aqueous medium in DMSO There is no significant difference in brain concentrations of E ₂ Q ⁺ . However, the latter produced significantly less variation and more consistent data. Interestingly, E ₂ Q ⁺ concentration in the lungs, is lower after administration of E ₂ -CDS-HPCD. The explanation for this, when administered in dissolved E ₂ -CDS in a water-miscible organic solvent such as DMSO, the
This very prone to water-insoluble E ₂ -CDS to precipitate considered or not located in some. After bolus intravenous injection, the ionic aqueous environment in the lungs may provide a suitable site for this deposition. E ₂ -CDS-HPCD is low density obtained in the lung after administration not only reflects the higher water solubility of the clathrate compound, also shows some characteristics of the in vivo dissociation constant May be. A completely unexpectedly, the rate of the dissociation, E ₂ -CDS in the CNS
Seems to be fast enough not to change the distribution and slow enough to pass through the lungs (or through organs such as the liver) without significant precipitation. Furthermore, the value of concentration in various organs also less variation much better after administration of E ₂ -CDS-HPCD. This can also be explained by the high water solubility of this clathrate compound and its low tendency to precipitate. Pharmacological experiments performed simultaneously also confirmed the efficacy of the E ₂ -CDS-HPCD composition for selective supply to the brain.

50% w / w solution of HPCD is used for many steroids and other CDS
The effect on the solubility of is shown in Table 2 below.

Thus, although E ₂ -CDS and a number of other CDS were successfully solubilized with HPCD, this is not a universal, for example, norethindrone -CDS is easy not solubilized. Maximum solubilization of E ₂ -CDS occurred in an aqueous solution of HPCD (ASDS 5.1 or 7). These clathrates could be freeze dried and were stable. This dry powder,
When the cyclodextrin component was 20% w / v or more, it was easily returned to a solution state by dissolving in water. Aqueous solution of this inclusion compound is E ₂ Q ⁺ the effect of supplying to the rat brain, E ₂ -
This was equivalent to the case where CDS was administered as a DMSO solution. In addition, this composition significantly reduced the lung concentration of E ₂ Q ⁺ . Data obtained to date indicate that this excipient (HPC
D) has been shown to be low toxic, easily compressible into tablets, rapidly dissolves, easy to synthesize and reproducible.

E ₂ is -CDS and other CDS addition, as described herein, could be formed even better inclusion compound using other cyclodextrin derivatives. Other cyclodextrin / CDS clathrates also exhibit improved properties as described above for HPCD clathrates.

In one example of a solid clathrate composition, a representative CD
The E ₂ -CDS is S, it was obtained from Ensuiko Sugar Co., Ltd.,
Excess to a 40% aqueous solution of a mixture of maltosyl-β-cyclodextrin (71%) of lot number 88190 and various dimaltosyl-β-cyclodextrins (29%),
Mix for several hours. The resulting suspension was then filtered, lyophilized and the amount of drug that formed the inclusion compound was determined by UV spectrophotometric analysis. The measured value of the degree of contamination is 81.88 mg / g
Met. The corresponding value for HPCD is about 36.2 mg / g.

In yet another test, the maximum concentration of each CDS that could be dissolved in aqueous solutions of various concentrations of each cyclodextrin was determined. C
The maximum concentration (mg / ml) when DS is E ₂ -CDS is shown below. Cyclodextrin 20% 40% HPCD 6.73 11.9 HEBCD 3.09 6.07 HEGCD-4.0 Norethindrone-CDS concentration (mg / ml) is 40% HPCD
7.13 in 40% HPGCD and 15.9 in HPGCD.

As described above, by using the E ₂ -CDS a representative dihydropyridine redox system drugs as inclusion compounds of a typical cyclodextrin derivative HPCD, the redox state of solution in DMSO The initial lung concentration of the quaternary form is lower (and therefore the initial brain / pulmonary concentration ratio is higher) than when the systemic drug is administered. Experiments with another representative redox drug, testosterone-CDS (T-CDS ₁ ), also gave similar experimental results, as described in detail below.

Experimental Procedure Materials Used 2-Hydroxypropyl-β-cyclodextrin (HPCD, degree of substitution 5.1) was prepared and purified according to the method of Pitha et al. Cyclodextrin inclusion compounds were prepared by mixing an excess of either testosterone propionate or T-CDS _{1 with} a 50% w / v aqueous solution of 2-hydroxypropyl-β-cyclodextrin to equilibrate. After degassing the solution and sonicating the suspension for 30 minutes, it was filtered and the filtrate was lyophilized. The dried filtrate is per gram of cyclodextrin clathrate.
65.6mg testosterone propionate or 29.6
It was found to contain a T-CDS ₁ of mg. Compounds were analyzed for degradation by thin layer chromatography and ultraviolet absorption.

Animals Male Sprague-Dawley rats weighing 250-275 g were
s Purchased from River Breeding Laboratories (Wilmington, MA) and housed in animals with controlled lighting (14 hours, 0500 hours light on) and temperature (23 ± 1 ° C.). Serum luteinizing hormone (LH)
To increase endogenous testosterone supply
Under light ether anesthesia, the right and left testes were removed by midline abdominal incision. All experiments began two weeks after this orchiectomy.

Experiment 1 On the 15th day after orchiectomy, the rats were anesthetized with ether to expose the right external jugular vein. The animals were then administered any of the following: testosterone-CDS (T-CDS ₁ or T-CDS
-CDS ₂ ), testosterone (from Steraloids Inc., Wilton, NH), or the excipient dimethyl sulfoxide (DMSO, from Fisher Scientific, Fairrain, NJ). Testosterone-CDS
Was administered to give a dose equal to testosterone (25 mg / kg). That is, T-CDS ₁ is 36.5 mg / kg,
-CDS ₂ was administered to rats in an amount of 45.1 mg / kg. DMSO 1ml
/ kg was injected. All compounds were administered by a 2 minute infusion. Immediately before drug administration (1000 hours), 1 ml of blood was collected from the external jugular vein, and blood samples were collected by cardiac puncture at 6, 12, 24 hours, and on days 4 and 7. The serum was separated by centrifugation at 500 xg for 20 minutes at 4 ° C and stored at -20 ° C.

Experiment 2 Two weeks after orchiectomy, T-CDS ₁ , testosterone propionate (TP, Steraloids Inc.) or DMS was used to more effectively increase the supply of testosterone to the brain.
Rats were administered either O by intravenous infusion into the right external jugular vein. Slow infusion has been shown to improve the delivery of drugs bound to this chemical delivery system to the brain. TP was selected for comparison because, like any T-CDS compound, it had an ester group (propionate group) bonded to the carbon at position ₁₇ (C ₁₇ ). Animals that were not orchidectomized received only drug vehicle. Two Harvard Apparatus reciprocating infusion / withdrawal pumps (model 944) were used so that four animals could be infused simultaneously. The injection rate is 15μ / min,
Infusion of the animals required 17-25 minutes. TP dose is 25mg
a / kg, T-CDS was injected in equimolar amounts of doses and TP (T-CDS ₁ 29.7mg of body weight 1 kg). The vehicle for this drug, DMSO, was administered at a dose of 1 ml / kg. 1 ml of blood is collected from the external jugular vein before infusion of the drug, and
And on day 7 from the suborbital sinus. Separation and storage of the serum were performed as described above.

Experiment 3 Testosterone-CDS (T-CDS ₁ ) (T-CDS ₁ -HPCD) dissolved in HPCD, testosterone propionade (TP-HPCD) dissolved in cyclodextrin, or excipient Were administered by a single tail vein injection. The dose of T-CDS ₁ -HPCD (11.9 mg / kg) was TP-H
The T-CDS ₁ dosage (10 mg TP / kg body weight) with an equimolar amount of the PCD decided to receive animals. Control rats were 3.0 ml /
kg of 25% HPCD (w / v) was administered. Blood was collected by cardiac puncture on days 0, 1, 3, 5, and 7, and separation and storage was performed as described above.

To evaluate the peripheral effects of the drug, the right seminal vesicle, vas deferens and ventral prostate are removed, cleaned, drained, and
Weighed to unit. The measured value is the weight (m
g).

LH radioimmunoassay (radioimmunoassay) Serum LH concentrations were measured twice with a tagioimmunoassay kit (reference formulation LH-RP-2) provided by the NIADDK pituitary hormone distribution program. The intra- and inter-assay coefficients of variation were 2.9 and 15.6, respectively.

Testosterone Radioimmunoassay Serum testosterone levels were measured using a coat A count (Cort
-A-Count) Radioimmunoassay Kit (Diagnos
Itic Products, Los Angeles, CA).

Statistical processing The significance of the difference between the mean values of LH and peripheral tissues was determined by analysis of variance (ANOVA) and Student-Newman-Keuls (SNK) tests. The significance level is
It was 0.05.

Results and Discussion In experiments 1 and 2 where DMSO was used as a vehicle for the drug, there were signs of insolubility of the drug after injection, that is, respiratory disturbances associated with lung lesions regardless of the rate of injection or infusion. To increase the water solubility of the steroids was dissolved Experiment 3 In T-CDS ₁ and TP in HPCD. T-
The improved solubility of CDS ₁ allowed lower doses (10 mg / kg vs. 25 mg / kg) to be administered, possibly reducing the risk of toxicity to animals.
Even if the dose of T-CDS ₁ 2.5 times less, resulting in suppression of serum LH levels comparable to that observed in the two experiments above (Experiment 1 and 2). Injection of T-CDS ₁ -HPCD resulted in a 50% decrease in serum LH by 24 hours, a suppression effect of 3%.
Allowed for days. In animals treated with TP-HPCD, the inhibitory effect of LH occurred only on day 1.

Mild stimulation of seminal vesicles by T-CDS ₁ -HPCD and T-
Mild stimulation of the ventral prostate with CDS ₁ -HPCD and TP-HPCD was observed 7 days after injection. As previously noted, the degree of stimulation by T-CDS ₁ -HPCD or TP-HPCD was slight relative to the tissue weight observed in control rats (without gonads removed). .

_One day after administration of T-CDS ₁ -HPCD to rats, a 5.5-fold increase in serum testosterone concentration was observed, and the serum testosterone concentration remained high until day 3. However, testosterone levels returned to pre-injection levels 5 days after injection. With TP-HPCD and HPCD, no increase in serum testosterone concentration occurred at any time point.

Above experiments are representative cyclodextrin T-CDS ₁ is a typical redox carrier drug HPCD
When the inclusion compound is used, the supply of testosterone to the brain is improved. In the experimental data, if T-CDS ₁ is an inclusion compound with HPCD, T-CDS ₁
Shows an equivalent suppression of LH even if the effective single dose of is reduced 2.5-fold. This finding, dihydropyridine form of T-CDS ₁ is an aqueous medium (e.g., blood) remains longer in solution state in, and thereby imply that passage of the blood brain barrier of the drug is improved. In previously conducted experiments, administration of DMSO as an excipient, in order to become deposited in the T-CDS ₁ probably blood (and Hi-Chew), apparent that cause respiratory failure and / or death in rats Has been. If the result of the administration of T-CDS ₁ in the form of inclusion compounds of HPCD did not occur death disordered breathing and animals. Obtained by a typical cyclodextrin HPCD, valleys initial lung concentrations of redox carrier compounds to quantify the effect of improving the effect of reducing as compared with the concentration in the brain, the HPCD inclusion compound of E ₂ -CDS Another series of experiments was performed using In this experiment detailed below, using reverse phase high performance liquid chromatography (HPLC) method, quaternary metabolite E its oxidized form and E ₂ -CDS in and tissues in a biological fluid ₂ - Quat was analyzed. This assay utilizes a precolumn enrichment technique, 10 ng / ml of E ₂ -Quat
And it detects a plasma concentration of up to E ₂ -CDS of 20 ng / ml. Preparation of a specimen (sample) is a quick and easy method. The sample is first homogenized with acetonitrile, centrifuged, and the supernatant is injected directly into the HPLC device.
The sample is injected into the pre-column by the water supply pump, where the drug is concentrated. The mobile phase allows the retained compounds to flow back to the analytical column. At the same time, another analyte can be injected into another pre-column. By this alternate pre-column sample concentration method, a large amount of injection up to 1800μ can be performed.

Experimental Operation Using material E ₂ -CDS, E ₂ -Quat and E ₂ -CDS-HPCD were synthesized as described above. Steroids (estradiol and ethinylestradiol) were obtained from Sigma Chemical Co. The mobile phase was prepared using acetonitrile for HPCD and distilled deionized water. All other reagents used were of analytical quality.

Instruments used HPLC equipment is LDC / Milton Roy Constametric III high pressure pump, LDC / Milton Roy variable wavelength UVUV detector, 2000μ
Perkin-Elmer ISS-100 auto-injector with loop of
And a 15 cm x 4.6 mm ID DuPont Zorbax ODS column (6 μm particle size). DuPont Zorba
x Vydac guard column dry-filled with ODS (5 cm x inside diameter
3.2 mm). The chromatogram is from Hewlett-Pack
A ard 3390A computer integrator was recorded at a chart speed of 0.2 cm / min. Furthermore, in this pre-column concentration device, a concentration injector (Rheodyne 7067-005) equipped with two high-pressure switching valves that are switched by air pressure by a tandem actuator (Rheodyne 7163) is connected to an automatic injector and an analytical column. Interposed between them. Switching of the valves was controlled by the automatic injector described above. The instrument also used a Bodine Electric Co. RP / 035 HP
An LC solvent pump was also provided.

Analytical Methods Analysis conditions: determined directly online HPLC E ₂ -CDS, the chromatographic conditions of analysis of E ₂ -Quat and estradiol (E _2). Optimum wavelength of all the compounds were 224nm but, E ₂ -CDS can be detected for dihydropyridine structure even 360 nm. The height of the absorbance at this wavelength is only about half the height of the absorbance at 224nm, but since the selectivity becomes high, was chosen 360nm as analytical wavelength of E ₂ -CDS. Various analytical columns were tested, and for all reversed phase chromatography of the three compounds, the mobile phase varied widely with respect to the ratio of aqueous: organic phase and buffer concentration and pH. No isocratic system could be found that could detect all three compounds with satisfactory peak widths and resolutions within a certain reasonable retention time. Therefore, two separate systems were used for the analysis.

E ₂ -Quat and E ₂ : The optimal mobile phase is acetonitrile / water containing 0.03 M sodium salt of octanesulfonic acid and 0.003 M tetrabutylammonium phosphate.
It was found to consist of a 0:60 mixture. The pH was adjusted to 5-5.5. The flow rate was 1.5 ml / min and the peak was recorded at 224 nm.

E ₂ -CDS: The mobile phase used for analysis of E ₂ -CDS, the flow rate 1.5
It was a 70:30 mixture of acetonitrile / water at ml / min.
Absorbance was monitored at 360 nm.

Analysis of E ₂ , E ₂ -CDS and E ₂ -Quat by pre-column enrichment: Sensitivity caused by the dilution process (see section on sample preparation without extraction), which is the optimal procedure for pretreatment of biological samples. Can be compensated for by developing an HPLC system that allows for large injections. Reference [Roth et al., J. Chro
matogr. 222 : 13-22 (1981)].
The method is a method based on alternate pre-column sample concentration. The method adopted in this experiment is as follows: the sample containing the drug is injected into one of the two pre-columns by means of the first pump A which supplies pure water. The two precolumns are alternately connected to the injector by two pneumatically driven valves. With some lipophilicity, the drug is retained and concentrated on this pre-column,
Water-soluble coproducts, such as entrained proteins, are washed away by the pump as long as the water is passing through the precolumn. This makes it possible to inject body fluid directly. After a certain concentration time (6 or 8 minutes), 2
The pre-column 1 which has caused the simultaneous rotation of the valves to absorb the injected drug is switched to the solvent flow of the second pump B. At this point, the recording integrator is started. Pump B supplies the mobile phase required for separation and chromatography, and allows the sample from precolumn 1 to flow back to the analytical column. In parallel with this operation, the pre-column 2 is switched to the water flow of the pump A so that the injection and concentration of the sample can be performed, and at the same time, the preceding pre-column performs elution (alternating mode). Due to the concentration effect of the concentrated phase, volumes up to 1800μ can be injected.

Chromatographic conditions: This instrument is compatible with E ₂ , E ₂ -CDS
And it could be applied to the quantification of E ₂ -Quat. Mobile phase E ₂ -CDS was a 80:20 mixture of acetonitrile / water at a flow rate 1.8 ml / min. The optimum peak shapes and retention times of E ₂ and E ₂ -Quat were as follows.
Obtained when feeding a 42:58 mixture of acetonitrile / water containing 0.003M /. Adjust the pH to 5 and the flow rate
It was 1.5 ml / min.

E ₂ -CDS containing standard solutions and stability respective 50μg / ml, E ₂ -Quat, a stock solution of the sample E ₂ and ethinyl -E ₂ was prepared by acetonitrile solvent. All solutions were stored at 6 ° C. E ₂ −CDS
For, the stock solution was freshly prepared every two weeks. All other solutions were stable for a period of at least 6 months.
Samples of spiked plasma containing all four compounds were frozen at -20 ° C and repeated at different time intervals. No drug loss was observed for 2 months under these storage conditions.

Dihydropyridine derivatives such as E ₂ -CDS are known to be easily oxidized in acidic solutions and very unstable. The stability of the E ₂ -CDS was investigated under various conditions at room temperature. This experiment, various pH stock solution of E ₂ -CDS
Diluted with various solvents or solutions at a 1: 2 ratio and monitored by monitoring the final peak height drop over 24 hours using the following variation of the direct online HPLC method described above . As changes, replace the water in the mobile phase of E ₂ -CDS in 0.05M phosphate buffer pH 7, 224n detection wavelength
If set to m, E ₂ -Quat it can be detected simultaneously at 6.33 min. However, this peak is relatively broad. Using this condition, to determine the degree of oxidation of E ₂ -CDS under the test conditions.

Sample Preparation E ₂ -Quat and E ₂ extraction: the various extraction from plasma were investigated under different conditions and several solvents and solvent mixtures. Estrone could be used as an internal standard, which is known to be a potential metabolite of estradiol. Therefore, 17β-ethynyl estradiol was chosen as the internal standard. Its peak did not interfere with E _2, E ₂ -Quat or estrone. Without the addition of the anion reagent, E ₂ -Quat could not be extracted from the aqueous solution. Optimum results were obtained after a one-step extraction using potassium iodide as the ion-pairing reagent to facilitate extraction of the quaternary salt. The method applied was as follows. 200 μ of a saturated potassium iodide solution was added to 1 ml of spiked plasma. After swirling and stirring for several seconds, 10 ml of a mixed solution of chloroform / ethyl acetate 9: 1 was added. After shaking the tube for 10 minutes,
Centrifuged at 00 rpm for 10 minutes. The upper aqueous phase was discarded and the organic layer was transferred to a clean test tube to achieve complete separation of protein and traces of the aqueous phase. Organic phase then under nitrogen
Evaporated to dryness at 40 ° C. and returned to solution with 150 μm mobile phase. 40μ was injected into the HPLC instrument. Appropriate blanks were prepared as described above.

E ₂ -CDS extraction: Plasma and water containing E ₂ -CDS,
Extraction was repeated with various organic solvents such as chloroform, hexane, toluene, benzene and ethyl acetate. However, extracting with good reproducibility E ₂ -CDS from the aqueous phase is not possible since this compound showed reproducible without deterioration during the evaporation even under a nitrogen atmosphere at room temperature and anoxia there were. Therefore, this compound can be obtained without performing an extraction operation.
It had to be analyzed from body fluids.

Extracted performed without the E ₂ -CDS and E ₂ -Quat and preparation of plasma and tissues for analysis of: by precolumn enrichment described above
The HPLC method can be used to detect drugs from directly injected plasma without sample preparation. However, if a large amount is injected to increase the sensitivity, it is desirable to remove a considerable amount of protein before injection in order to prevent the pre-column from being frequently packed. Subsequent E ₂ -CDS so that only one sample preparation process is required
And we were selected protein removal method which can be applied to the analysis of both E ₂ -Quat.

Small amount capable of removing proteins with only added to the body fluid acidic deproteinization (precipitation) agent, could not be used because it causes loss due to decomposition of E ₂ -CDS.
Neutral to weakly basic aqueous reagents used efficiently for protein removal, such as ZnSO ₄ / NaOH, CuSO ₄ / Na ₂ SO ₄ , or saturated (NH ₄ ) ₂ SO _4, are more concentrated than organic solvents. It would be more ideal to inject into the activated column. However, none of these reagents were shown to absorb the E ₂ -CDS water-insoluble adhered to precipitate. So avoid instability issues,
The method chosen to keep all compounds in solution at the same time was protein removal with acetonitrile.

To obtain these results, the following sample preparation procedure was used on plasma and tissues. Plasma: 0.6 ml of plasma 1.2 m
Added to 1 liter of acetonitrile. The mixture was swirled and stirred for 5 seconds, left at room temperature for 10 minutes, swirled and stirred again,
Centrifuged at 2000 rpm for 10 minutes. 1000-1500μ of the supernatant was injected into the pre-column concentrator. Tissue (eg, brain): 1
Add ml of water to one rat brain and mesh the organs thoroughly. Sonicate for 2 minutes, 10 at 2000 rpm
After centrifugation for 1 minute, the supernatant (1000-1500μ) is injected into the concentration device.

In animal experiments a first experiment, the E ₂ -CDS of 15 mg / kg dissolved in dimethylsulfoxide (DMSO), administered intravenously constrained male Sprague-Dawley rats conscious of the weight 190~300g did. Animals divided into groups of 4 were given 5, 15 and 30 minutes after injection of the drug and 1, 2, 4, 8,
Dissected 24 and 48 hours later. Trunk blood was collected in heparinized tubes and plasma was obtained and immediately frozen at -20 ° C until analysis. Organs were excised within 2 minutes of death and placed on dry ice and stored at -20 ° C for later analysis by the HPLC method described above.

In the second experiment, the same operation was performed as above,
However, 5 mg / kg of E ₂ -CDS was administered in the form of an aqueous solution as an inclusion compound with hydroxypropyl-β-cyclodextrin (HPCD). An amount of 5 mg / kg of E ₂ -CDS is added to about 20% w / v
In 1 ml of an aqueous solution containing This aqueous solution is 3.5
prepared by dissolving clathrate lyophilized compound containing E ₂ -CDS in mg / g (prepared from solution of E ₂ -CDS 50% aqueous solution of HPCD degree of substitution 5.1) in 20% HPCD solution did.

Results and Discussion The results are shown in FIG. 1 and FIG. Figure 1 is the concentration of E ₂ -CDS in lung tissue (graph on the left) and E ₂ -Quat (right graph), corrected for dose as μg amount (C _B / D) per dose 1g And a pair of plots on a semilog scale. From FIG. 1, it can be seen that when E ₂ -CDS is administered as a solution in DMSO, the initial lung concentration (ie, the concentration within the first hour after drug injection) is equal to that of E ₂ -CDS and E ₂ -Quat. both, E ₂ -CDS significantly compared to the initial lung concentrations observed when administered in aqueous form as inclusion compounds of HPCD (10 times or more) higher were it can be seen.
(Correction corner also dose) is the concentration of E ₂ -Quat corresponding in brain tissue, the dose 1g at a particular point in time in FIG. 2
It is shown in the form of a bar graph showing the brain concentration (C _B / D) as the ng amount per unit. It can be seen that the concentration in the brain 1 hour after administration was not significantly different when administered in the form of an aqueous solution as an HPCD inclusion compound, compared with administration in the form of a DMSO solution. Accordingly, the carrier-drug can be administered in the form of an aqueous solution as an inclusion compound with a specific cyclodextrin, such as HPCD, used in the present invention, in which case the desired biological effect (pharmacological action) can still be obtained. Obviously, it is possible to obtain the necessary brain concentrations to produce, while at the same time avoiding the high initial lung concentrations that cause dyspnea and dysnia.

The formation of inclusion compounds with 2-hydroxypropyl-β-cyclodextrin (HPCD) and other cyclodextrin derivatives used in the present invention has been found to be particularly advantageous in stabilizing dihydropyridine redox systems. Direct comparison of the stability in an aqueous solution form, a low solubility in water of the dihydropyridine-type redox system drugs (e.g., solubility in water of E ₂ -CDS is only 0.0002 mg / ml) for, of course impossible. E ₂ -CDS-HPCD inclusion compound containing E ₂ -CDS about 40mg to 1g, easily formed into 1ml cyclodextrin concentration of 20% w / v aqueous solution containing E ₂ -CDS of 5mg I do. Therefore, formation of inclusion compounds, resulting 25,000-fold increase in water solubility of E ₂ -CDS. The half-life of E ₂ -CDS in aqueous solution when placed in the dark at room temperature is about 12 5 days. (Speed: 0.0554 ± 0.0047d ^-1).

Since dihydropyridine redox drugs are particularly susceptible to deterioration due to oxidation, an experiment was conducted to quantify the effect of cyclodextrin derivatives such as HPCD used in the present invention on the oxidation rates of these drugs. Typical carriers - the E ₂ -CDS are agents were used in this experiment.

The rate of ferricyanide-mediated oxidation of E ₂ -CDS was determined by previously published methods (Okamoto et al ., J. Chem . Soc . Comm .,
1977, 181). In this experimental operation, 5 ×
The 10 ^-3 M concentration of E ₂ -CDS in acetonitrile 27.5Myu,
1 × 10 ^-4 M Fe (CN) ₆ ^-4 , 0.06 M K ⁺ , 0.001 M Fe (C
Was added to a solution 2.75ml containing N) ₆ ^-3. All solutions were prepared using water that was boiled for 30 minutes and then cooled while passing a stream of nitrogen scrubbed with pyrogallol. E ₂ -CDS was introduced at 37 ° C. in a thermostatted cell holder into a solution contained in an anaerobic screw-top cuvette from a syringe.
The cuvette had a septum lined with Teflon and the compound was injected through the septum. Constant concentration of ferricyanide ion (6 × 10 ^-4 to 8 × 10 ^-3 M)
Was measured for the disappearance rate of E ₂ -CDS. This was done by calculating the decrease in the absorbance band at 360 nm (± 10 nm) subtracted from the baseline absorbance (500 ± 10 nm). The slope of the pseudo-first-order rate constant was obtained by plotting 1n (absorbance) against time. This was done at several different ferricyanide ion concentrations. The obtained first-order rate constant was then plotted against the ferricyanide ion concentration, and the second-order rate constant (k ₀ s ⁻¹ M ⁻¹ ) was determined from the obtained slope. In examining the effect of 2-hydroxypropyl-β-cyclodextrin on the oxidation rate of E ₂ -CDS, a solution containing HPCD and each ion present in the above first-stage experiment was prepared. Second order rate constants were determined for each cyclodextrin concentration and plotted. The results obtained show that the presence of cycloextrin drastically reduces the oxidation rate. When the cyclodextrin concentration exceeds 2% w / v, no significant change in the oxidation rate is observed and it appears that there is a saturation effect. The secondary oxidation rate was suppressed by 42% with 0.5% w / v cyclodextrin, 60% with 1.0% w / v cyclodextrin, 81% with 2% w / v cyclodextrin, and a concentration of 5 to 20%. In w / v, about 90% reduction of the oxidation rate was obtained.

From the above, by preparing or formulating a drug composition using a representative cyclodextrin derivative used in the present invention, a lipid-reduced form of a dihydropyridine pyridinium salt-type redox carrier system for drug delivery targeted to the brain It is clear that the problems associated with the administration of are overcome. Specifically, the above-mentioned specific cyclodextrin derivative, preferably hydroxypropyl-β-
When a carrier / drug composition for parenteral administration in the form of an aqueous solution containing cyclodextrin at a concentration of about 20 to 50% w / w or w / v is administered, the distribution of the drug unexpectedly changes,
It has been found that the problem of drug deposition in the lungs associated with the use of organic solvents is avoided and reduced toxicity is obtained. Also,
The time required for the drug to separate from the cyclodextrin after injection is long enough to prevent coagulation of the drug molecule (ie, drug coagulate deposition) in the lungs or other organs (eg, liver). However, the favorable temporal conditions are such that the drug / cyclodextrin degradation is short enough to provide easy distribution of the drug molecule to achieve the desired pharmacological action. It was unexpected.

Was not previously available for parenteral administration at all?
Alternatively, other lipophilic / hydrophobic and / or water-labile drugs that could not be formulated for parenteral administration without the use of organic solvents, to varying degrees, were found in the redox carrier system. However, such other drugs may benefit from the same improved drug distribution and favorable time conditions as described above. Particularly useful agents for use in the pharmaceutical compositions and methods for parenteral administration of the present invention are certain cyclodextrin derivatives (eg, HPCD) that are relatively insoluble in water but have a concentration of 20-50% in water. Can be substantially increased by co-prescribing the compound. Such properties can be determined by simple experiments of the type described below for representative drugs.

Equipment Ultraviolet spectra were recorded on a Cary 210 double beam spectrophotometer (Varian). Micromeritics 728 autosampler, Beckmann 112 solvent delivery module, Waters Lambd
a-Max 481 LC spectrophotometer and Fisher Recordall 50
HPLC (High Pressure Liquid Chromatography) was performed on a device having a configuration of a 00 series recording device. Samples were from Fisher Brans
Sonicated with an onic ultrasonic cleaner and temperature equilibrated in a MGW Lauda constant temperature water bath.

Solubility experiment Add excess test drug to aqueous solution containing 2-hydroxypropyl-β-cyclodextrin in various amounts,
A phase solubility experiment was performed by sonicating the mixture for 1 hour. After equilibration for at least 48 hours in a water bath at 25 ± 1 ° C. in the dark, a part of the mixture is removed.
The solution was filtered through a 0.45 μm membrane filter, diluted, and the drug concentration was measured by a reverse phase HPLC method.

 The solubility of the drug in water was also measured for comparison.

HPLC method Chlordiazepoxide Wavelength: 245 nm Column: Waters μBondapak CN, inner diameter 3.9 mm × 30 cm Mobile phase: acetonitrile / acetic acid / water containing 0.1% 1-hexanesulfonic acid sodium salt (60: 1: 39) Flow rate 2.00 ml / min Retention time: 4.0 minutes Dexamethasone wavelength: 263 nm Column: ASI C18, 10 μm, inner diameter 3.9 mm × 30 cm Mobile phase: acetonitrile / water (55:45) Flow rate: 1.00 ml / min; Storage time: 3.6 minutes Diazepam wavelength: 241 nm column : Waters μBondapak CN, inner diameter 3.9 mm × 30 cm Mobile phase: acetonitrile / water (6: 4) Flow rate: 2.00 ml / min; retention time 3.2 minutes 17β-estradiol Wavelength: 280 nm Column: ASI C18, 10 μm, inner diameter 3.9 mm × 30 cm Mobile phase: acetonitrile / water (55:45) flow rate: 2.00 ml / min; storage time: 4.4 minutes 17α-ethynyl estradiol wavelength: 248 nm column: Fisher Resolvex C18, inner diameter 4.6 mm × 25 cm mobile phase: acetonitrile / water (6: 4) Flow rate: 1.50ml / min; Retention time: 4.4 minutes Estradiol 3-methyl ether Wavelength: 248 nm Column: Fisher Resolvex C18, inner diameter 4.6 mm × 25 cm Mobile phase: acetonitrile / water (7: 3) Flow rate: 2.00 ml / min; Storage time: 6.0 minutes Medazepam Wavelength: 253 nm Column: Waters μBondapak CN, inner diameter 3.9 mm x 30 cm Mobile phase: acetonitrile / acetic acid / water (0.1: 39) containing 0.1% 1-hexanesulfonic acid sodium salt Flow rate: 2.00 ml / min; Retention time: 2.8 minutes Methotrexate Wavelength: 308 nm column: Fisher Resolvex C18, 4.6 mm ID x 25 cm mobile phase: methanol / acetic acid / water (50: 1: 49) containing 0.1% 1-octanesulfonic acid sodium salt Flow rate: 2.00 ml / min; retention time: 3.5 min Norethindrone Wavelength: 240 nm Column: Fisher Resolvex C18, 4.6 mm id x 25 cm Mobile phase: Acetonitrile / water (6: 4) Flow rate: 1.50 ml / min; Retention time: 3.6 min Norethindrone acetate Wavelength: 240 nm Column: Fisher Resolvex C18 , 4.6mm inside diameter × 25cm Mobile phase: Ase Nitrile / water (7: 3) Flow rate: 2.00 ml / min; Storage time: 5.0 minutes D (-)-norgestrel Wavelength: 241 nm Column: Fisher Resolvex C18, 4.6 mm ID x 25 cm Mobile phase: acetonitrile / water (7: 3) ) Flow rate: 2.00 ml / min; Storage time: 3.4 minutes Oxazepam wavelength: 230 nm Column: Waters μBondapak CN, inner diameter 3.9 mm × 30 cm Mobile phase: acetonitrile / water (35:65) Flow rate: 2.00 ml / min; Retention time 2.6 minutes Phenytoin Wavelength: 258 nm Column: Fisher Resolvex C18, 4.6 mm id x 25 cm Mobile phase: acetonitrile / water (55:45) Flow rate: 2.00 ml / min; Storage time: 1.6 minutes All-trans-retinol Wavelength: 325 nm Column: Waters μBondapak C18, inner diameter 3.9 mm x 30 cm Mobile phase: acetonitrile / water (55:45) Flow rate: 2.00 ml / min; retention time 5.8 minutes TABLE 4 Increased solubility of drug by 25% w / v aqueous solution of hydroxypropyl-β-cyclodextrin at 25 ± 1 ° C. Drug solubility (mg / ml) Dexamethasone 24.18 17β-estradiol 19.13 17α-ethynylestradiol 34.47 17α-ethynylestradiol 3-Methyl ether 7.13 Norethindrone 9.13 Norethindrone acetate 9.41 D (-)-norgestrel 2.19 instrument Ultraviolet spectra were measured on a Perkin-Elmer 550 SE double beam spectrophotometer. Rheodyne 7125 injector, LKB 2150 HPLC pump, LKB 2138 Lichrosorb RP 1
High pressure liquid chromatography was performed on an apparatus configured with an 8 10 mm column (4 x 250 mm), an LKB 2138 Uvicord 5 detector and a 0mniscribe recorder. The samples were sonicated in a Kerry ultrasonic bath and equilibrated in a Tecam TE-Tempette constant temperature water bath.

Solubility experiments An excess of the test agent is added to an aqueous solution containing various amounts of 2-hydroxypropyl-β-cyclodextrin (HPCD) and the mixture is sonicated for up to 4 hours to determine the phase solubility. An experiment was performed. In the dark
After equilibration in a water bath at 30 ± 0.2 ° C. for up to 72 hours, a portion of the above mixture was filtered through a 0.45 μm membrane filter, diluted, and the drug concentration was measured by HPLC or UV. Due to drug instability, sonication and equilibration times were minimized.

HPLC method Chlorambutyl Wavelength: 245 nm Mobile phase: acetonitrile / acetic acid / water (45: 1: 54) Flow rate: 2.00 ml / min Retention time: 4.4 minutes Lomustine wavelength: 254 nm Mobile phase: methanol / water (7: 3) Flow rate: 2.00 ml / min Retention time: 3.6 minutes Melphalan Wavelength: 254 nm Mobile phase: methanol / acetic acid / water (60: 1: 39) + 0.19% 1
-Pentanesulfonic acid sodium salt Flow rate: 2.00 ml / min Retention time: 3.6 minutes Result Chlorambutyl Solubility of approx. 30 mg / g in 50% w / w aqueous solution of HPCD (30 min. 4 hours equilibration treatment). This drug is almost insoluble in water,
The oral dose is about 100 mg / kg / day. The results obtained from further experiments are shown in Table 5 below. Significant degradation occurred during the experiment (3.5 days).

Lomustine Initial experiments showed a solubility of about 12 mg / g in 50% w / w aqueous HPCD. The results of the follow-up experiment are shown in Table 6 below.
Some decomposition occurred.

Melphalan In the first experiment, about 21.9mg / g in 50% w / w HPCD aqueous solution
(After sonication for 1 hour, equilibration treatment at 30 ° C. for 4 hours). The results of the tracking experiment are shown in Table 7 below.
Some decomposition occurred.

Table 6 Solubility of lomustine in aqueous solution of HPCD at 30.0 ± 0.2 ° C HPCD concentration (% w / v) Solubility (mg / ml) ^* 0 0.18 1 0.38 5 1.68 10 3.33 15 6.26 20 8.44 25 8.9 *) Is the average of up to four experiments Table 7 : Solubility of melphalan in aqueous solution of HPCD at 30.0 ± 0.2 ° C HPCD concentration (% w / v) Solubility (mg / ml) ^* 0 1.26 1 4.16 2 4.3 3 6.24 4 7.14 5 7.2 7 10.5 10 13.37 15 17.64 20 24.75 25 31.36 *) Values shown are the average of several experiments Solubility experiments Phase solubility experiments are carried out by adding an excess of alphaxalone to a phosphate buffer containing a specific cyclodextrin derivative at a concentration of 20% w / v and sonicating the mixture for at least 1 hour. Was done. (This phosphate buffer dissolves 1.183 g of KH ₂ PO ₄ and 4.320 g of Na ₂ HPO ₄ in water,
Prepared by diluting to 1. ) The mixture was filtered through a 0.45 μm Millipore syringe filter, diluted and the drug concentration was determined by reverse phase HPLC. Alphaxalone itself is substantially insoluble in water.

Alphaxalone HPLC method Wavelength: 294 nm Mobile phase: methanol / water (65:35) Flow rate: 1.5 ml / min Retention time: 1414 minutes The results of this experiment are summarized in Table 9 below.

TABLE 9 Solubility of alphaxalone in 20% w / v solution of various cyclodextrin derivatives at room temperature Cyclodextrin solubility (mg / ml) Maltosyl-β-cyclodextrin mixture 20.02 Hydroxypropyl-γ-cyclodextrin 18.41 Hydroxypropyl- γ-cyclodextrin 15.11 Hydroxypropyl-β-cyclodextrin 22.83 Hydroxypropyl-β -cyclodextrin 24.42 The following examples target a brain that can be used in combination with a cyclodextrin derivative (eg, HPCD) according to the present invention. This is an example of a method for synthesizing a preferred specific example of a reduced form (dihydropyridine compound) of a dihydropyridine pyridium salt-type redox carrier system for supplying a drug. These have not been specifically described in publications so far.

Example 1 Synthesis of N-nicotinoyl dopamine 11.7 g (0.05 mol) of dopamine hydrobromide and nicotinic acid
To a pyridine solution at 0 ° C. containing 6.15 g (0.05 mol),
10.3 g of dicyclohexylcarbodiimide (DCC) (0.05mo
l) was added. The reaction mixture was stirred at room temperature for 24 hours, and by-produced sidiclohexylurea was removed by filtration. The pyridine was removed under reduced pressure and the residue was crystallized from water at 0 ° C.
The product was filtered off and dried over phosphorus pentoxide. Recrystallization from isopropanol gave 9.0 g (0.035 mol) of N-nicotinoyldopamine (70%). Melting point 159-162
° C. Aqueous solution of this compound ^turns green due to Fe ³⁺ , Ag
NO ₃ was reduced.

IR (KBr): 3300,2960,1725,1630,1590,1520,1430,1290,
1190, 1115, 720, 710 cm ^-1 ; NMR (d ₆ -DMSO) δ 9.25-6.25 (m, 7H), 3, 3 (m, 2H), 2.
65 (m, 2H) ppm; Elemental analysis: matching _{C 14 H 14 N 2 O 3} .

Example 2 Synthesis of 1-methyl-3- {N- [β- (3,4-dihydroxyphenyl) ethyl]} carbamoylpyridinium iodide 2 g (7.7 mol) of N-nicotinoyldopamine dissolved in 40 ml of dry methanol To 2.5g of methyl iodide (17.6mm
ol) was added. The reaction mixture was heated at reflux with stirring for 6 hours. An additional 1.5 g (1.05 mmol) of methyl iodide was added and reflux continued overnight. The methanol was removed and ethyl acetate was added to give the desired product as yellow crystals. Yield 2.
4g (77%). 173-174 ° C.

Example 3 Synthesis of 1-methyl-3- {N- [β- [3,4-bis (isobutyryloxy) phenyl] ethyl]} carbamoylpyridinium trifluoroacetate 3 g (7.5 mmol) of the product of Example 2 ) Was dissolved in 30 ml of trifluoroacetic acid in an ice-cooled solution, and isobutyryl chloride 2.
4 g (22.5 mmol) were added slowly with stirring. Stirring was continued at room temperature overnight. The trifluoroacetic acid was evaporated under reduced pressure and the residue was crystallized from ethyl ether / hexane (3: 1). Yield 1.2 g (30.4%). 87-91 ° C.

Example 4 1-methyl-3- {N- [β- [3,4-bis (isobutyryloxy) phenyl] ethyl]} carbamoyl-1,4
Synthesis of dihydropyridine 1-methyl-3- {N- [β- [3,4-bis (isobutyryloxy) phenyl] ethyl]} carbamoylpyridinium in 50 ml of degassed water containing 10 ml of methanol
A solution of 0.55 g (1 mmol) of trifluoroacetate was extracted with three 30 ml portions of ether. To the resulting aqueous solution was added 0.25 g (3 mmol) of NaHCO ₃ and 50 ml of ethyl ether, and the mixture was kept under a nitrogen atmosphere. To this ice-cooled mixture was added 0.52 g (3 mmol) of sodium dithionite and the mixture was stirred vigorously for 30 minutes. The ether layer was separated and the aqueous layer was extracted twice with ether. The combined ether extracts were washed and dried over sodium sulfate. The ether was removed in vacuo leaving an oily product. NMR analysis confirmed that the product had the structural formula shown below.

Example 5 Synthesis of 5,5-diphenyl-3-hydroxymethyl-2,4-imidazolidinedione 5 g (0.02 mol) of phenytoin was suspended in 180 ml of water,
It was added formaldehyde (37% aqueous solution) 20 ml and K ₂ CO ₃ 0.25g, and the resulting mixture was stirred for 24 hours at 25 to 30 ° C.. The precipitated white solid was separated by filtration, washed several times with a 3% aqueous formaldehyde solution, air-dried for 3 to 4 hours, dried in a desiccator under reduced pressure, and dried over P ₂ O ₅ . Yield 91-93%. Melting point 185-1
89 ° C.

Elemental analysis (as C ₁₆ H ₁₄ O ₃₎ Calculated: C, 68.07; H, 5.00 ; N, 9.93 Found: C, 67.97; H, 5.05 ; N, 9.93.

 This product is represented by the following formula.

Example 6 Synthesis of 5,5-diphenyl-3-[(3'-pyridyl) carbonyloxymethyl] -2,4-imidazolidinedione 3.00 g (0.011 mol) of the product of Example 5 was added to dry pyridine 1
Dissolve in 50 ml and then 4.25 g (0.019 mol) of nicotinic anhydride
Was added. The resulting solution is allowed to dry at room temperature (25-30
C) for 40 hours. This solution was poured into water of 2.5, and the precipitated white solid was separated by filtration, washed well with water, placed in a desiccator under reduced pressure, and dried over P ₂ O ₅ . 95% yield. Melting point
178-182 ° C.

Elemental analysis (C ₂₂ H ₁₇ N as ₂ O ₄₎ Calculated: C, 68.21; H, 4.42 ; N, 10.85 Found: C, 68.12; H, 4.43 ; N, 10.83.

 This product is represented by the following formula.

Example 7 Synthesis of 5,5-diphenyl-3-[(1'-methyl-3'-pyridinium) carbonyloxymethyl] -2,4-imidazolidinedione iodide 0.5 g (0.0013 mol) of the product of Example 6 ) Is dissolved in 50 ml of acetonitrile, then 0.3 ml of methyl iodide is added,
The resulting reaction mixture was kept at room temperature for 6 days. The solvent was distilled off under reduced pressure, and ethyl ether was added to the residue. After cooling the ether solution for 2 hours in a refrigerator, the precipitated yellow hygroscopic crystals were placed in a vacuum desiccator and dried over P ₂ O ₅ to give the desired product in 85% yield. The UV and H ¹ NMR spectra confirmed that the obtained product was represented by the following structural formula.

The above reaction operation, using nitromethane as a solvent,
Use a bath temperature of 50-70 ° C and use excess methyl iodide for 5
Repeated by gradually adding in ６6 hours gave the same product in almost quantitative yield.

Example 8 5,5-diphenyl-3-[(1'-methyl-1 ', 4'-
Synthesis of dihydropyridin-3'-yl) carbonyloxymethyl] -2,4-imidazolidinedione 0.4 g (0.0008 mol) of the quaternary salt obtained in Example 7 was dissolved in 40 m of water.
l, a mixed solvent consisting of 3 ml of methanol and 15 ml of ethyl acetate. After cooling the reaction mixture to 0-5 ° C and degassing, 0.39 g (0.0046 mol) of sodium bicarbonate and 0.54 g (0.0032 mol) of sodium dithionite were added. The mixture was stirred at 0-5 ° C for 35 minutes under a nitrogen atmosphere. The organic layer was separated and the aqueous layer was extracted with two 15 ml portions of ethyl acetate. These organic solutions were extracted with 10 ml of cold degassed water. After drying over sodium sulfate, the solvent was distilled off under reduced pressure, and the residual oily yellow solid was crystallized by adding ether. yield
70%. The UV and H ¹ NMR spectra confirmed that the obtained product was represented by the following structural formula.

Example 9 3-bromoacetyloxymethyl-5,5-diphenyl-
Synthesis of 2,4-imidazolidinedione HCl was generated in 2 g (0.0071 mol) of 5,5-diphenyl-3-hydroxymethyl-2,4-imidazolidinedione in 15 g (8 ml, 0.096 mol) of bromoacetyl chloride. Dissolution was achieved by heating in an oil bath (bath temperature 70-80 ° C) for about 15 minutes until stopped. The resulting mixture was cooled and 30 ml of ethyl ether was added. White crystals precipitated. After cooling the mixture to 0 ° C., the crystals were filtered off and dried over P ₂ O ₅ . Yield 2.15 g (75%). Melting point 179-183 [deg.] C.

Elemental analysis _{(C 18 H 15 N 2 O} 4 as Br) Calculated: C, 53.61; H, 3.75 ; N, 6.95; Br, 19.82 Found: C, 53.60; H, 3.79 ; N, 6.92; Br, 19.90 This product is represented by the following formula.

Example 10 3- (3'-bromopropionyl) oxymethyl-5,5
Synthesis of -diphenyl-2,4-imidazolidinedione According to the method described in Example 9, 5,5-diphenyl-
3-hydroxymethyl-2,4-imidazolidinedione 5 g
(0.018 mol) was reacted with 6.8 g (0.04 mol, 4 ml) of 3-bromopropionyl chloride using an oil bath temperature of 100 ° C. 4.9 g (65% yield) of the product as white crystals were obtained. 133-134 ° C.

Elemental analysis (as C ₁₉ H ₁₇ N ₂ O ₄ Br) Calculated: C, 54.69; H, 4.11; N, 6.72; Br, 19.15 Found: C, 54.79; H, 4.12; N, 6.69; Br, 19.25 This product is represented by the following formula.

Example 11 3- (2'-bromopropionyl) oxymethyl-5,5
Synthesis of diphenyl-2,4-imidazolidinedione 2 g (0.0071 mol) of 5,5-diphenyl-3-hydroxymethyl-2,4-imidazolidinedione was 8.5 g (5 ml, 0.05 mol) of 2-bromopropionyl chloride Inside, 100-110
Dissolution was achieved by heating in a 30 ° C. oil bath for 30 minutes. The reaction mixture was cooled, 20 ml of ethyl ether was added, and the resulting solution was extracted with an aqueous potassium carbonate solution, dried, and crystallized. The product was obtained as a white solid (1 g, 3%). 112-115 ° C.

Elemental analysis (as C ₁₉ H ₁₇ N ₂ O ₄ Br) Calculated: C, 54.69; H, 4.11; N, 6.72; Br, 19.15 Found: C, 54.77; H, 4.15; N, 6.69; Br, 19.25 This product is represented by the following formula.

Example 12 Synthesis of 3- (3'-carbamoyl-1'-pyridinium) acetyloxymethyl-5,5-diphenyl-2,4-imidazolidinedione bromide The product of Example 9 dissolved in 15 ml of nitromethane 2.02
g (0.005 mol) was mixed with nicotinamide (0.61 g, 0.005 mol). The resulting solution was stirred for 2 hours in an oil bath at a bath temperature of 90-100 ° C. The mixture was cooled to 60 to 70 ° C., and the precipitated white crystals were separated by filtration and washed with nitromethane. yield
61% (1.65 g).

193-197 ° C (decomposition).

Elemental analysis (C ₂₄ H ₂₁ N ₄ O as ₅ Br) Calculated: C, 54.87; H, 4.03 ; N, 10.67; Br, 15.21 Found: C, 54.70; H, 4.05 ; N, 10.64; Br, 15.25 This product is represented by the following formula.

Example 13 3- [3 '-(3 "-carbamoyl-1" -pyridinium) propionyloxymethyl] -5,5-diphenyl-
Synthesis of 2,4-imidazolidinedione bromide 2.09 g (0.005 mol) of the product of Example 10 is dissolved in 15 ml of acetonitrile and then 0.61 g (0.005 g) of nicotinamide
mol) was added. After the resulting solution was heated under reflux for 6 days, the solvent was removed. To the resulting gummy residue was added 30 ml of ethyl ether and the mixture was stirred for 2 hours. The precipitated white substance was separated by filtration and washed with ether. Yield 78% (2.
1g). 98-100 ° C (decomposition). UV and H ¹ NMR spectra were as expected. This product is represented by the following formula.

Example 14 3- [2 '-(3 "-carbamoyl-1" -pyridinium) propionyloxymethyl] -5,5-diphenyl-
Synthesis of 2,4-imidazolidinedione bromide 0.69 g (0.00165 mol) of the product of Example 11 is dissolved in 8 ml of acetonitrile and then 0.2 g (0.0016 g) of nicotinamide
5 mol) was added and the resulting solution was heated at reflux for 22 hours.
The solvent was removed from the resulting brown amorphous material at 50 ° C., then 15 ml of ethyl ether were added and the mixture was stirred for 2 hours. The obtained light brown substance was separated by filtration and washed with ether. Yield 56% (0.5 g). 158 ° C (decomposition). This product is represented by the following formula.

Example 15 Synthesis of 3-[(3'-carbamoyl-1 ', 4'-dihydropyridin-1'-yl) acetyloxymethyl] -5,5-diphenyl-2,4-imidazolidinedione Preparation of Example 12 0.52 g (0.001 mol) of the product was dissolved in a mixture of 60 ml of water and 30 ml of ethyl acetate. The solution was cooled to 5 ° C. and degassed, then 0.5 g of sodium bicarbonate (0.006 mol
l) and 0.7 g (0.004 mol) of sodium dithionite were added, and the resulting mixture was stirred for 30 minutes while being degassed and cooled. After liquid separation, the aqueous layer was extracted with 30 ml of ethyl acetate. The organic solution was extracted with 20 ml of cooled and degassed water. After drying over sodium sulfate, the solvent was removed. Yellow crystals with a melting point of 155 DEG-160 DEG C. (decomposition) were obtained in a 55% yield (0.25 g), which reduced an alcoholic solution of silver nitrate and had the formula

Example 16 3- [3 '-(3 "-carbamoyl-1", 4 "-dihydropyridin-1" -yl) propionyloxymethyl]
Synthesis of -5,5-diphenyl-2,4-imidazolidinedione Following the general reaction procedure described in Example 15,
The reduction with sodium dithionite using the product of 13 was repeated essentially as in Example 15, yielding the desired product in 85% yield. The melting point of this product is 10
It was 0 ° C (decomposition) and represented by the following structural formula.

The product of Example 14 was similarly reduced and converted to the corresponding dihydro derivative. 105 ° C (decomposition).

Example 17 Synthesis of 4-aminobutyric acid cyclohexyl ester hydrochloride 8 g (77.6 mmol) of GABA was suspended in 100 ml (0.96 mmol) of cyclohexanol. Add thionyl chloride 40 to this mixture.
ml was added dropwise at 0 ° C. The resulting mixture was heated at reflux for 4 hours, then cooled and crystallized from ethyl ether. The white crystals thus obtained were filtered off and dried. NMR analysis confirmed the title product.

Example 18 Synthesis of 3- {N-[(3'-cyclohexyloxycarbonyl) propyl]} carbamoylpyridine 2.2 g (18 mmol) of nicotinic acid was suspended in 50 ml of dry pyridine. Dicyclohexylcarbodiimide is added to this suspension.
3.68 g (17.9 mmol) were dissolved with stirring. 4 g (18 mmol) of 4-aminobutyric acid cyclohexyl ester hydrochloride were added and the mixture was stirred for 48 hours. The precipitated dicyclohexylurea was removed by filtration, and the filtrate was evaporated to dryness. 25 ml of residue
Was washed with ice-cold water and extracted into ethyl acetate. Liquid separation,
The organic layer was evaporated to dryness. NMR analysis confirmed the structure of the title product.

Example 19 Synthesis of 1-methyl-3- {N '-[(3'-cyclohexyloxycarbonyl) propyl]} carbamoylpyridinium iodide 1.74 g (6 mmol) of the product of Example 18 was dissolved in a minimum amount of acetone. , And the precipitated white precipitate was removed by filtration. Methyl iodide (1.5 ml, 24 mmol) while stirring at 0 ° C. in the resulting solution
Was added all at once. The mixture was gently refluxed overnight. The white precipitate was filtered off and the remaining yellow filtrate was evaporated, leaving a reddish oil. This was dissolved in acetone, filtered and evaporated to dryness.

Elemental analysis (as C ₂₂ H ₂₃ N ₂ O ₃ I) Calculated: C, 47.26; H, 5.79; N, 6.48; I, 29.38 Found: C, 47.03; H, 5.85; N, 6.44; I, 29.26 Example 20 Synthesis of 1-methyl-3- {N-[(3'-cyclohexyloxycarbonyl) propyl]} carbamoyl-1,4-dihydropyridine 0.11 g (0.26 mmol) of the product of Example 19 was cooled in 25 ml of ice Dissolved in degassed water. NaHCO ₃ 0.09 g (4-fold excess) followed by Na ₂
0.14 g (3 fold excess) of S ₂ O ₄ was added. 25 ml of ethyl acetate were added and the mixture was stirred for 30 minutes under a nitrogen atmosphere. The organic layer was extracted and dried to give an orange oil. This immediately reduced the methanol solution of silver nitrate. NMR analysis confirmed that the product had the following structure.

Example 21 Synthesis of valpro acid chloride (2-propylpentanoyl chloride) Thionyl chloride was added to 4.32 g (30 mmol) of valproic acid in an ice bath.
3.60 g (30 mmol) were added slowly with stirring. The solvent-free mixture is allowed to warm to room temperature and then in a 50 ° C water bath.
Heat for 30 minutes. Add 50 ml of dry benzene twice each,
Removed under reduced pressure. The obtained product was used for the next reaction without further purification.

Example 22 Synthesis of valproic acid 2-iodoethyl ester (2'-iodoethyl-2-propylpentanoate) 4.87 g (30 mmol) of the product of Example 21 was added thereto while cooling and stirring in an ice bath. -5.16 g of iodine ethanol
(30 mmol) was added. This solventless mixture is then added to 10
After heating in a water bath at 0 ° C. for 10 minutes, it was removed from the water bath and stirred for another 10 minutes. The reaction mixture was then dissolved in 50 ml of ether and washed with water (1 × 30 ml), 5% NaOH (2 × 30 ml), and again with water (2 × 30 ml). The ether layer was dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. 67% yield (6.0 g) of pale yellow liquid product from valproic acid
Was obtained. Silver nitrate produced a bright yellow precipitate. NM
The product was confirmed to be the title product by R analysis.

Example 23 1- [2 '-(2 "-propyl) pentanoyloxy]
Synthesis of ethyl-3-carbamoylpyridinium iodide The product of Example 22 in 1.22 g (10 mmol) of nicotinamide
3.28 g (11 mmol) and 50 ml of dimethylformamide were added. The mixture was heated at reflux for 3 hours and then cooled.
Removal of the solvent under reduced pressure gave a brown oily residue. This was stirred with 60 ml of ether for 30 minutes to give a yellow powder. The ether was decanted off and 50 ml of fresh ether was added. The crude product was filtered under reduced pressure under a nitrogen atmosphere, and then recrystallized from isopropanol / ether to give the desired product.
3.5 g (84% yield) was obtained. 111-112 ° C. This product was represented by the following structural formula.

Example 24 1- [2 '-(2 "-propyl) pentanoyloxy]
Synthesis of ethyl-3-carbamoyl-1,4-dihydropyridine The product 42 of Example 23 was added to 50 ml of ice-cold degassed deionized water.
0 mg (1 mmol) was added. NaHCO ₃ 366mg to the resulting solution
(4 mmol) and 696 mg (4 mmol) of Na ₂ S ₂ O ₄ were added with stirring. Nitrogen gas was bubbled through the solution for 30 minutes. The aqueous solution was then extracted with ether (6 × 25 ml) until the ether layer was no longer yellow. The combined ether extracts were washed with water (1 × 50 ml) and dried over magnesium sulfate. The ether layer was decanted from the desiccant and the solvent was removed under reduced pressure. After ether was added to the obtained oily residue, it was removed by a vacuum pump (10 × 5 m
l). Bubbles evolved, which returned to oil upon exposure to the atmosphere. NMR analysis confirmed the structure.

Example 25 Synthesis of N-nicotinoyl tyrosine ethyl ester 12.3 g (0.1 mol) of nicotinic acid was dissolved in 300 ml of dry pyridine. The solution was cooled and 20.6 g (0.1 mol) of dicyclohexylcarbodiimide was added. After dissolution, 24.6 g (0.1 mol) of tyrosine ethyl ester hydrochloride was added,
The resulting solution was stirred overnight. The precipitated dicyclohexylurea (DCU) was removed by filtration. The resulting oil was triturated with boiling water to further remove DCU. This product was purified with acetone.

Elemental analysis _{(C 17 H 18 Na 2 O} 4 · 1 / 2H 2 O ) Calculated values: C, 63.16; H, 5.88 ; N, 8.66 Found: C, 63.10; H, 5.96 ; N, 8.59 The product Is N- [1-ethoxycarbonyl-2-
(4'-Hydroxyphenyl) ethyl] nicotinamide.

Example 26 N-[(1-methyl-3-pyridinium) carbonyl]
Synthesis of tyrosine ethyl ester iodide N-nicotinoyl tyrosine ethyl ester 20g (0.06m
ol) was dissolved in 200 ml of acetone. A 2-fold molar excess of methyl iodide (25.6 g, 0.18 mol) was added and the mixture was added to 6 parts.
Heated to reflux for hours. The solvent was removed under reduced pressure to give the desired product as a solid. NMR analysis confirmed that it was the title compound represented by the following structural formula.

This compound can also be named 1-methyl-3- {N-[(1'-ethoxycarbonyl) -2 '-(4 "-hydroxyphenyl) ethyl} carbamoylpyridinium iodide.

Example 27 Synthesis of 1-methyl-3- {N-[(1'-ethoxycarbonyl) -2 '-(4 "-pivaloyloxyphenyl) ethyl]} carbamoylpyridinium trifluoroacetate Preparation of Example 26 6 g (0.013 mol) of the product were dissolved in 50 ml of cold trifluoroacetic acid at 0 ° C. in an ice bath.
g (0.026 mol) was added slowly and the solution was allowed to warm to room temperature. After 24 hours, the solvent was removed under reduced pressure. The resulting dark oil was triturated with petroleum ether, but no solid precipitated. The identity of the product was confirmed by NMR analysis. This product was dissolved in aqueous methanol (10%) and extracted with ethyl ether to remove highly colored contaminants before being used as starting material in Example 29 below.

Example 28 Synthesis of 1-methyl-3- {N-[(1'-ethoxycarbonyl) -2 '-(4 "-isobutyloxyphenyl) ethyl]} carbamoylpyridinium trifluoroacetate 6g of product of Example 26 (0.013 mol) was dissolved in 50 ml of trifluoroacetic acid in an ice bath cooled to 0 ° C. To this solution, 2.77 g (2.76 ml) of isobutyryl chloride was gradually added while stirring. The mixture was stirred overnight, and the solvent was removed under reduced pressure.The resulting oil was stirred with petroleum ether overnight and dried under reduced pressure, but no solid precipitated, and the identity of the product was confirmed by NMR analysis. This product was dissolved in aqueous methanol (10%) and extracted with ethyl ether to remove highly colored contaminants.
Used as starting material in Example 30 below.

Example 29 Synthesis of 1-methyl-3- {N-[(1'-ethoxycarbonyl) -2 '-(4 "-pivaloyloxyphenyl) ethyl]} carbamoyl-1,4-dihydropyridine 4.07 g (0.0079 mol) of the product was dissolved in 10 ml of a 25% aqueous methanol solution, nitrogen gas was bubbled through the solution, and the solution was stirred in an ice bath while NaHCO ₃ 2.
02 g (0.024 mol) were added. Ethyl ether 100ml was added, followed by addition of _{Na 2 S 2 O 4 4.12g (} 0.024mol). The obtained yellow two-phase solution was stirred for 30 minutes, then separated, and the aqueous layer was separated for 75 minutes.
Extracted twice with each ml of ethyl ether. The combined organic extracts were dried over sodium sulfate and the solvent was removed under reduced pressure to give a solid foam. It was oxidized by ethanolic silver nitrate solution.

Elemental analysis (as C ₂₃ H ₂₀ Na ₂ O ₅ .1 / 2H ₂ O) Calculated: C, 65.23; H, 7.33; N, Found: C, 65.76; H, 7.28; N, 6.95 Synthesis of -methyl-3- {N-[(1'-ethoxycarbonyl) -2 '-(4 "-isobutyryloxyphenyl) ethyl]} carbamoyl-1,4-dihydropyridine 2.20 g of the product of Example 28 the (0.0044 mol) was dissolved in aqueous methanol solution 100 ml. the solution was cooled in an ice bath while a stream of nitrogen gas thereto. NaHCO ₃ 1.11 to this solution
g (0.0132 mol) and 100 ml of ether were added. Then 2.30 g (0.0132 mol) of sodium dithionite were added and the solution was stirred for 30 minutes. After liquid separation, the aqueous layer was washed with ethyl ether. The combined organic layers were dried over anhydrous Na ₂ SO _4, and concentrated. The resulting orange oil was oxidized with a silver nitrate ethanol solution. The identity of the product was confirmed by NMR analysis.

Example 31 Chloromethyl. [2S- (2α, 5α, 6β)]-3,3-dimethyl-7-oxo-6-[(2,6-dimethoxy) benzamide] -4-thia-1-azabicyclo [3.2 .0] Synthesis of heptane-2-carboxylate To a solution of 4.02 g (0.01 mol) of methicillin sodium salt dissolved in 10 ml of water and 10 ml of methylene chloride, 2.4 g of sodium bicarbonate and 0.34 g of tetrabutylammonium hydrogen sulfate were added. did. Then, 1.9 g (0.0115 mol) of chloromethyl chlorosulfate dissolved in 3 ml of methylene chloride was added with stirring over 5 minutes while maintaining the temperature at 30 ° C or lower. After stirring for an additional 30 minutes, the organic phase was separated, washed twice with water and dried over MgSO _4. Removal of the solvent under reduced pressure gave 4.24 g of the desired product as a yellow solid, mp 88-90 ° C.

Example 32 Chloromethyl. [2S- (2α, 5α, 6β)]-3,3-dimethyl-6- (5-methyl-3-phenyl-4-isoxazolecarboxamide] -7-oxo-4-thia- 1
Synthesis of azabicyclo [3.2.0] heptane-2-carboxylate 2.12 g (0.005 mol) of oxacillin sodium salt was treated with NaH
CO ₃ 1.2 g, used with tetrabutylammonium hydrogen sulfate 0.17g, and chlorosulfonic acid chloromethyl 0.95 g, by repeating substantially the reaction procedures described in Example 31,
1.87 g of the expected product with a melting point of 78 DEG-80 DEG C. (decomposition) are obtained.

Example 33 Chloromethyl. [2S- (2α, 5α, 6β)]-6- [3-
(2-Chlorophenyl) -5-methyl-4-isoxazolecarboxamide] -3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2
Synthesis of carboxylate By the same reaction procedure as in Example 31 except that cloxacillin sodium salt 2.38 g (0.005 mol) was used as the reactant.
(1 mol water), 1.2 g of NaHCO ₃ , 0.17 g of tetrabutylammonium hydrogen sulfate and 0.95 g of chloromethyl chlorosulfate gave 2.27 g of the desired product with a melting point of 97-100 ° C. (decomposition).

Example 34 Chloromethyl. [2S- (2α, 5α, 6β)]-6- [3-
Synthesis of (2,6-dichlorophenyl) -5-methyl-4-isoxazolecarboxamide] -3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylate In accordance with the method of Example 31 except that 2.55 g (0.005 mol) of dicloxacillin sodium salt is used as a reactant.
(1 mol water), 1.7 g of NaHCO ₃ , 0.17 g of tetrabutylammonium hydrogen sulfate and 0.95 g of chloromethyl chlorosulfate gave 2.43 g of the desired product with a melting point of 98-101 ° C. (decomposition).

Example 35 [(3-pyridinylcarbonyl) oxy] methyl • [2S
-(2α, 5α, 6β)]-3,3-dimethyl-7-oxo-
Synthesis of 6-[(2,6-dimethoxy) benzamide) -4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylate 3.8 g of methicillin chloromethyl ester obtained in Example 31 in 70 ml of dimethylformamide (0.0089 mol) and 1.6 g (0.01 mol) of potassium nicotinate at room temperature (20-25 ° C)
For 6 days. 300 ml of ethyl acetate was added, and the precipitated solid was separated by filtration.
Extracted once and dried over MgSO ₄ . The solvent was removed under reduced pressure,
The obtained residue was purified by chromatography (silica gel). 3 g of the expected product with a melting point of 151-157 ° C. were obtained as a white solid.

Example 36 [(3-pyridinylcarbonyl) oxy] methyl • [2S
-(2α, 5α, 6β)]-3,3-dimethyl-6- (5-methyl-3-phenyl-4-isoxazolecarboxamide) -7-oxo-4-thia-1-azabicyclo [3.
2.0] Synthesis of heptane-2-carboxylate According to the method of Example 35, but with 1.81 g (0.004 mol) of oxacillin chloromethyl ester obtained in Example 32
Is reacted with 0.75 g (0.0046 mol) of potassium nicotinate to obtain, after purification by chromatography, 0.1 g of the desired product.
75 g was obtained as a white solid, mp 79-82 ° C (decomposition).

Example 37 [(3-pyridinylcarbonyl) oxy] methyl • [2S
-(2α, 5α, 6β)]-6- [3- (2-chlorophenyl) -5-methyl-4-isoxazolecarboxamide] -3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [ 3.2.0] Synthesis of heptane-2-carboxylate Utilizing the method of Example 35, but using 2.1 g of the cloxacillin chloromethyl ester obtained in Example 33 (0.0043 mol
l) and 0.8 g (0.005 mol) of potassium nicotinate gave 1.2 g of product with a melting point of 83-85 ° C. (decomposition).

Example 38 [(3-pyridinylcarbonyl) oxy] methyl • [2S
-(2α, 5α, 6β)]-6- [3- (2,6-dichlorophenyl) -5-methyl-4-isoxazolecarboxamide] -3,3-dimethyl-7-oxo-4-thia-1
Synthesis of azabicyclo [3.2.0] heptane-2-carboxylate Similarly, using the method of Example 35, except that 2.27 g of the dicloxacillin chloromethyl ester obtained in Example 34
(0.0047 mol) and 0.87 g of potassium nicotinate (0.0054mo)
The reaction of l) gave 11 g of the expected product as a white solid with a melting point of 87-90 ° C. (decomposition).

Example 39 [2S- (2α, 5α, 6β)]-3-[[[[[3,3-dimethyl-7-oxo-6-[(2,6-dimethoxy) benzamide] -4-thia-1 Synthesis of -Azabicyclo [3.2.0] hept-2-yl] carbonyl] oxy] methoxy] carbonyl] -1-methylpyridinium iodide 1.25 g (0.0024 mol) of the methicillin derivative obtained in Example 35 in 35 ml of nitromethane 1.14 g of methyl iodide (0.5m
l) (0.008 mol) was reacted for 7 days in a closed apparatus at room temperature (20-25 ° C). The solvent was removed under reduced pressure and the remaining residue was stirred with ether, filtered, washed with ether and dried. A yellow, hygroscopic product having a melting point of 95-100 ° C. was thus obtained. This was confirmed to be represented by the following structure.

Example 40 [2S- (2α, 5α, 6β)]-3-[[[[[3,3-dimethyl6- (5-methyl-3-phenyl-4-isoxazolecarboxamide) -7-oxo-4 -Thia-1-
Synthesis of Azabicyclo [3.2.0] hept-2-yl] carbonyl] oxy] methoxy] carbonyl] -1-methylpyridinium iodide Using the method of Example 39, but reacting the material in Example 36 in 25 ml of nitromethane. Oxacillin derivative obtained in
0.5 g (0.0009 mol) and _{CH 3 I0.45g (0.2ml) (0.003mol} )
In place of, after 6 days, the melting point of 75 to
0.6 g of the expected product at 80 ° C. was obtained.

Example 41 [2S- (2α, 5α, 6β)]-3-[[[[[6- [3-
(2-Chlorophenyl) -5-methyl-4-isoxazolecarboxamide] -3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] hept-2-
Yl] carbonyl] oxy] methoxy] carbonyl]-
Synthesis of 1-methylpyridinium iodide Similarly, utilizing the method of Example 39, except that the cloxacillin derivative obtained in Example 37 in 25 ml of nitromethane
0.44 g (0.0008 mol) and CH _{3 I} 0.45 g (0.2 ml) (0.003 mol)
Is reacted, melting point 90 ~ 95 ℃ shown by the following structural formula
0.45 g of (decomposed) product was obtained.

Example 42 [2S- (2α, 5α, 6β)]-3-[[[[[6- [3-
(2,6-Dichlorophenyl) -5-methyl-4-isoxazolecarboxamide] -3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] hept-
Synthesis of 2-yl] carbonyl] oxy] methoxy] carbonyl] -1-methylpyridinium iodide Similarly, the procedure of Example 39 was used, except that 0.5 g of the dicloxacillin derivative obtained in Example 38 in 25 ml of nitromethane. (0.0007mol) and CH ₃ I0.45g (0.2ml) (0.003mo
l) to give a melting point of 95-1
0.55 g of product at 00 ° C. (decomposition) was obtained.

Example 43 [[(1,4-dihydro-1-methyl-3-pyridinyl)
[Carbonyl] oxy] methyl • [2S- (2α, 5α, 6
β)] Synthesis of -3,3-dimethyl-7-oxo-6-[(2,6-dimethoxy) benzamido] -4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylate Degassed 0.45 g (0.0007 mol) of the product of Example 39 dissolved in a mixed solvent of 25 ml of ethyl acetate and 70 ml of water was treated with NaHCO ₃ 0.34
g (0.004mol) and sodium dithionite 0.48g (0.0028mo
The mixture with l) was reduced at 0-5 <0> C for 70 minutes. Disappearance of the maximum absorption at 268 nm in the ultraviolet spectrum and 3
The increase in the maximum absorption at 66 nm was followed. After liquid separation, the aqueous layer was extracted with 2 × 25 ml of ethyl acetate, and the organic layer was extracted with 2 × 25 ml of ethyl acetate.
Extracted with 20 ml cold degassed water. The organic layer was dried with sodium sulfate, and the solvent was removed under reduced pressure. 0.25 g of product
Obtained as a yellow solid with a melting point of 88-90 ° C (decomposition). The product was of the formula:

Example 44 [[(1,4-dihydro-1-methyl-3-pyridinyl)
[Carbonyl] oxy] methyl • [2S- (2α, 5α, 6
β)]-3,3-Dimethyl-6- (5-methyl-3-phenyl-4-isoxazolecarboxamide) -7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylate The general reaction method of Example 43 was repeated except that Example 40 was carried out in a mixed solvent of 15 ml of water and 15 ml of ethyl acetate.
0.17 g (0.00025 ml) of NaHCO ₃ 0.08 g (0.0001mo
l) and 0.51 g (0.001 mol) of Na ₂ S ₂ O ₄ were reacted. 0.1 g of product with a melting point of 93-100 ° C. (decomposition) was obtained. This product had a structure represented by the following formula.

Example 45 [[(1,4-dihydro-1-methyl-3-pyridinyl)
[Carbonyl] oxy] methyl • [2S- (2α, 5α, 6
β)]-6- [3- (2-chlorophenyl) -5-methyl-4-isoxazolecarboxamide] -3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.
2.0] Synthesis of heptane-2-carboxylate Similarly, according to the method of Example 43, but using as reactants 0.18 g (0.00025 ml) of the product of Example 41, NaHCO ₃
Reaction of 0.089g and Na ₂ S ₂ O ₄ 0.17g, mp 80-85
0.13 g of the product as a yellow solid at ℃ (decomposition) were obtained. This product had a structure represented by the following formula.

Example 46 [[(1,4-dihydro-1-methyl-3-pyridinyl)
[Carbonyl] oxy] methyl • [2S- (2α, 5α, 6
β)]-6- [3- (2,6-dichlorophenyl) -5
Methyl-4-isoxazolecarboxamide] -3,3
Synthesis of -dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylate Similarly, 0.19 g (0.00025 ml) of the product of Example 42, NaHC
The procedure of Example 43 was repeated using 0.08 g of O ₃ and 0.17 g of Na ₂ S ₂ O ₄ to obtain 0.14 g of the desired product having a melting point of 98 to 102 ° C. (decomposition) and represented by the following structural formula. .

Example 47 [(3-pyridinylcarbonyl) oxy] methyl • [2S
-(2α, 5α, 6β)]-3,3-dimethyl-7-oxo-
6-[(phenylacetyl) amino] -4-thia-1-
Synthesis of azabicyclo [3.2.0] heptane-2-carboxylate The chloromethyl ester of benzylpenicillin in 100 ml of dimethylformamide, ie chloromethyl. [2S-
(2α, 5α, 6β)]-3,3-dimethyl-7-oxo-6
-[(Phenylacetyl) amino] -4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylate
3.83 g (0.01 mol) and potassium pyridine-3-carboxylate
A suspension with 1.93 g (0.012 mol) was stirred at 20-25 ° C. for 6 days. Then, 300 ml of ethyl acetate was added, and the precipitated solid was separated by filtration. The solution was extracted four times with a concentrated aqueous sodium chloride solution and dried over magnesium sulfate. Removal of the solvent under reduced pressure gave 4.5 g of a foamy solid. Purification by chromatography on silica gel with ethyl acetate as eluent gave 2.5 g of product with a melting point of 127-130 ° C.

Example 48 [2S- (2α, 5α, 6β)]-3 [[[[[3,3-dimethyl-7-oxo-6-[(phenylacetyl) amino]
-4-thia-1-azabicyclo [3.2.0] hept-2-
Yl] carbonyl] oxy] methoxy] carbonyl]-
Synthesis of 1-methylpyridinium iodide 2.5 g (0.053 mol) of the product of Example 47 dissolved in 100 ml of dry nitromethane was added to 2.25 g (1 ml) of methyl iodide (0.
016 mol) in a closed apparatus at 20 to 25 ° C for 6 days.
At the end of the reaction period, the reaction was complete based on the results of thin layer chromatography. The solvent was removed under reduced pressure and the solid residue was slurried with ether, filtered and dried over P ₂ O ₅ under reduced pressure. The product, mp 90-95 <0> C (decomposition) was obtained as a yellow solid (2.91 g). The following structural formula was assigned to this product.

Example 49 [[1,4-Dihydro-1-methyl-3-pyridinyl) carbonyl] oxy] methyl • [2S- (2α, 5α, 6β)]
-3,3-dimethyl-7-oxo-6-[(phenylacetyl) amino] -4-thia-1-azabicyclo [3.2.
0] Synthesis of heptane-2-carboxylate 3.25 g of quaternary salt compound prepared in Example 48 (0.0
053 mol) was dissolved in a mixed solvent of 350 ml of water and 150 ml of ethyl acetate. The resulting mixture was cooled to 0-5 ° C and degassed with nitrogen, and then a mixture of 2.67 g (0.032 mol) of sodium bicarbonate and 3.69 g (0.021 mol) of sodium dithionite was added to the mixture.
Added over 3 minutes. The reaction mixture was stirred for 1 hour under the same conditions as above, and then separated. The aqueous layer was extracted twice with 50 ml portions of ethyl acetate, and the combined organic extracts were washed twice with 30 ml portions of cold degassed water. The organic layer was dried over sodium sulfate and the solvent was removed under reduced pressure to obtain 1.7 g of a yellow solid product having the following structural formula and a melting point of 98 to 100 ° C.

Example 50: Chloromethyl N- [3- (10,11-dihydro-5H-dibenzo [b, f] azepin-5-yl)] propyl-N-
Synthesis of methyl carbamate Method A Desipramine hydrochloride (1.5 g, 0.005 mol) was added at 0-5 ° C.
And dissolved in 20 ml of methylene chloride. Then 1 g of NaHCO ₃ followed by 0.92 g of chloromethyl chloroformate
(0.007 mol) was added. After stirring the reaction mixture for 1 hour, the precipitated salts were filtered off and the solution was extracted twice with 10 ml portions of 5% HCl. The organic layer was dried over magnesium sulfate, and the solvent was removed under reduced pressure to obtain the target compound as a colorless oily substance in a yield of 76% (1.35 g).

Method B Imipramine hydrochloride (1.59 g, 0.005 mol) was dissolved in 15 ml of water and then 5 ml of a 4% aqueous sodium hydroxide solution was added with cooling. The resulting imipramine base was extracted twice with 10 ml portions of benzene. Concentrate the extract to 10 ml,
Then 5 ml of 0.7 g (0.0054 mol) of chloromethyl chloroformate
Of benzene dissolved in benzene was added while cooling to 10 ° C. The reaction mixture was stirred at 20-25 ° C. for 30 minutes and then heated at reflux for 1 hour. A small amount of imipramine hydrochloride formed and was filtered off. The resulting solution is diluted with 2 portions of 20 ml of 4% HCl.
Extracted once and dried over magnesium sulfate. Removal of the solvent under reduced pressure gave 1.2 g (66%) of the product with the same properties as obtained in Method A.

Example 51 1-chloroethyl.N- [3- (10,11-dihydro-5H
-Dibenzo [b, f] azepin-5-yl)] propyl-
Synthesis of N-methyl carbamate Following the general reaction procedure described in Example 50, but
Using 1.5 g (0.005 mol) of desipramine hydrochloride and 0.86 g (0.006 mol) of chloroethyl chloroformate, the reaction was carried out for 5 to 10 minutes.
1.6 g (86%) of the title compound as a colorless oil
I got

Example 52 [{N- [3- (10,11-dihydro-5H-dibenzo [b,
f] Synthesis of azepin-5-yl)] propyl-N-methylamino {carbonyloxy] methyl-3-pyridinecarboxylate 0.57 g (0.046 mol) of nicotinic acid and 0.4 of triethylamine
Example 5 in which 5 g was dissolved in 5 ml of dimethylformamide in a solution prepared by dissolving 5 g in 5 ml of dimethylformamide.
Product 0 (1.35 g, 0.0037 mol) was added. The resulting mixture was stirred at 25-30 ° C. for 24 hours, then ethyl acetate 30
ml was added. The precipitated salt was separated by filtration, and the obtained solution was 15 ml.
Extraction four times with saturated aqueous sodium chloride solution. Drying over magnesium sulfate and removal of the solvent under reduced pressure gave 1 g (61%) of the oily pure product.

Example 53 [1- {N- [3- (10,11-dihydro-5H-dibenzo [b, f] azepin-5-yl)] propyl-N-methylamino} carbonyloxy] ethyl-3-pyridinecarboxy Synthesis of the Rate Following the general reaction procedure described in Example 52, except that
1.05 g (0.0028 mol) of the product of Example 51, nicotinic acid 0.45
g (0.036 mol) and 0.36 g of triethylamine,
The reaction was performed in 10 ml of dimethylformamide at 25-30 ° C. for 48 hours to give 0.5 g of the title compound as a yellow oil.

Example 54 3-[{N- [3- (10,11-dihydro-5H-dibenzo [b, f] azepin-5-yl)] propyl-N-methylamino} carbonyloxy] methoxycarbonyl-1-
Synthesis of methylpyridinium iodide 0.8 g of the product of Example 52 in 30 ml of nitromethane (0.0018
mol) was reacted with methyl iodide (0.8 ml) at 25-30 ° C. for 48 hours. After the solvent was removed under reduced pressure, the residue was slurried form with ethyl ether, filtered and dried over P ₂ O _5. The title quaternary salt was obtained in 83% yield (0.88 g). The product is a pale yellow solid having a melting point of 172 to 175 ° C (decomposition) and is represented by the following structural formula.

Example 55 3- [1- {N- [3- (10,11-dihydro-5H-dibenzo [b, f] azepin-5-yl)] propyl-N-methylamino} carbonyloxy] ethoxycarbonyl-
Synthesis of 1-methylpyridinium iodide Generally following the alkylation procedure described in Example 54,
However, 0.5 g of the product of Example 53 in 15 ml of nitromethane (0.
0011 mol) and 0.5 ml of methyl iodide,
By running at 25 ° C. for 6 days, 0.33 g (50%) of the desired quaternary salt having the following structure was obtained as a dark yellow solid having a melting point of 101-103 ° C. (decomposition).

Example 56 [{N- [3- (10,11-dihydro-5H-dibenzo [b,
f] Azepin-5-yl)] propyl-N-methylamino {carbonyloxy] methyl-1,4-dihydro-1-
Synthesis of methyl-3-pyridine carboxylate 0.3 g (0.005 mol) of the product of Example 54 dissolved in a mixed solvent of 30 ml of water and 15 ml of ethyl acetate was dissolved in sodium bicarbonate at a concentration of 0.1 ml.
25g (0.003mol) and sodium dithionite 0.35g (0.002m
ol), a reduction treatment was carried out for 60 minutes while degassing at 0 to 5 ° C. After separation, the aqueous layer was extracted twice with 30 ml portions of ethyl acetate. The combined organic layers were then extracted twice with 20 ml portions of cold degassed water. The organic layer was dried over sodium sulfate and the solvent was removed under reduced pressure to give 0.22 g (95%) of the title compound having the following structural formula and a melting point of 59-63 ° C (decomposition).
was gotten.

Example 57 [1- {N- [3- (10,11-dihydro-5H-dibenzo [b, f] azepin-5-yl)] propyl-N-methylamino} carbonyloxy] ethyl-1,4- Dihydro-
Synthesis of 1-methyl-3-pyridinecarboxylate According to the method of Example 56, but with 10 ml of water and ethyl acetate
0.1 g (0.0017 mol) of the product of Example 55 in 6 ml, NaHCO ₃ 0.
The reaction is carried out for 60 minutes using 11 g and 0.15 g of Na ₂ S ₂ O ₄ to obtain 0.07 g (88%) of the title compound as a yellow solid having a melting point of 60 to 65 ° C. (decomposition) represented by the following structural formula. Was done.

Example 58 Synthesis of N- (2-hydroxyethyl) -3-pyridinecarboxamide A solution of 49.2 g (0.32525 mol) of ethyl nicotinate and 72 g (1.17 mol) of ethanolamine was heated to 70 ° C. for 60 hours. Excess ethanolamine was removed under reduced pressure and the resulting viscous creamy oil was stirred with ether for 48 hours. The precipitated white solid was filtered off to give 46 g of the title compound (8
5.1%). Melting point 75-78 [deg.] C Example 59 3- [2 '-(2 "-propyl) pentanoyloxy]
Synthesis of ethylcarbamoylpyridine 1.0 g (0.006021 mol) of the product of Example 58 and 0.61 g (0.00598 mol) of triethylamine were dried in 40 m of dichloromethane.
The solution dissolved in l was stirred, and to this solution, 1.96 g (0.012047 mol) of (2-propyl) pentanoyl chloride was added, and the mixture was heated under reflux for 4 hours. 5% of the resulting solution
Washed sequentially with 30 ml of NaHCO _3, 30 ml of 5% HCl and 30 ml of water. The organic layer was dried over MgSO _4, and the solvent was removed under reduced pressure, a pale product 0.6 g (34.3%) of oily brown was obtained.

Example 60 Synthesis of 1-methyl-3- [2 ′-(2 ″ -propyl) pentanoyloxy] ethylcarbamoylpyridinium iodide 1 g (0.00342 mol) of the product of Example 59 was dried with ethyl acetate
To a solution dissolved in 20 ml, add methyl iodide 0.73 g (0.00513mo
l) was added. The reaction mixture was stirred overnight at room temperature. The precipitated pale yellow solid was separated by filtration and recrystallized from ethyl acetate to give the title quaternary salt (1.35 g, 90.9%) as a yellow crystalline solid. This product has the formula:
Confirmed by IR, NMR, and UV analysis.

Example 61 Synthesis of 1-methyl-3- [2 '-(2 "-propyl) pentanoyloxy] ethylcarbamoyl-1,4-dihydropyridine 50 ml of degassed deionized water was vigorously stirred while cooling with ice, A solution of 3.0 g (0.006909 mol) of the quaternary product of Example 60 in 50 ml of ethyl acetate was added. Retained sodium bicarbonate 3.
5 g (0.04145 mol) and sodium dithionite 4.8 g (0.02
(763 mol). After 45 minutes, the organic layer was separated and the aqueous layer was extracted with 100 ml of ice-cooled ethyl acetate. The organic layer was washed with ice-cold water and the combined and dried over MgSO _4. Removal of the solvent under reduced pressure gives the product of the formula
2.1 g (98.8%) was obtained as a pale yellow solid.

The above structure was confirmed by IR, NMR and UV analysis.

Example 62 Synthesis of 3-pyridinecarboxylic acid (2-hydroxy) ethyl ester hydrochloride 16 ml of thionyl chloride was added dropwise to 120 ml of cold (−10 ° C.) ethylene glycol. After dropping, 24.6 g of nicotinic acid
(0.2 mol) was added little by little, and the resulting reaction mixture was added to 60
Heated to ° C overnight. Then, hot tetrahydrofuran 700m
l was added and the mixture was cooled. The precipitated solid was separated by filtration and washed with ether to obtain 28.5 g of the title compound as white crystals.

Example 63 3- [2 '-(2 "-propyl) pentanoyloxy]
Synthesis of ethoxycarbanylpyridine A solution of 10.0 g (0.0491 mol) of the product of Example 62 in 150 ml of dry methylene chloride, 10.7 g of triethylamine (0.
09819 mol) was added. After all solids were dissolved, 11.92 g (0.07364 mol) of 2-propylpentanoyl chloride was added and the reaction mixture was stirred at room temperature for 36 hours. The resulting solution was washed sequentially with 5% NaHCO ₃ , 5% HCl and water,
The organic layer was dried over anhydrous MgSO _4. Removal of the solvent under reduced pressure gave a tan oil which was triturated with a 40:60 mixture of ether and petroleum ether to yield 9.7 g of product as an orange oil.

Example 64 Synthesis of 1-methyl-3- [2 '-(2 "-propyl) pentanoyloxy] ethoxycarbonylpyridinium iodide 2.0 g (0.006816 mol) of the product of Example 63 was dissolved in 10 ml of dry acetone. 1.45 g of methyl iodide (0.01022mo
l) was added and the mixture was heated at reflux overnight. Removal of the solvent under reduced pressure provided 1.84 g of the title quaternary salt as a brown oil. This product is represented by the following formula.

Example 65 Synthesis of 1-methyl-3- [2 '-(2 "-propyl) pentanoyloxy] ethoxycarbonyl-1,4-dihydropyridine 50 ml of degassed deionized water was vigorously stirred while cooling with ice, A solution of 1.84 g (0.004226 mol) of the quaternary product of Example 64 in 50 ml of ethyl acetate was added during the reaction, while the temperature and pH were kept at 0 ° C. and 8 respectively, while bubbling argon through the reaction mixture. NaHCO ₃ 2.13 g
(0.02536mol) and was a mixture of _{Na 2 S 2 O 4 2.94g (} 0.0169mol) was added in portions. After 55 minutes, the organic layer was separated and the aqueous layer was extracted with 100 ml of ice-cooled ethyl acetate. The organic layer was washed with ice-cold water and the combined and dried over MgSO _4. Removal of the solvent under reduced pressure gave 0.9 g of the title compound as a yellow oil. This product was represented by the following formula.

Example 66 3,17β-bis [(3-pyridinylcarbonyl) oxy]
Synthesis of -19-nor-17α-pregna-1,3,5 (10) -trien-20-yne To 2.0 g (6.7 mmol) of ethinylestradiol dissolved in 50 ml of dry pyridine was added 6.16 g (0.027 mol) of nicotinic anhydride.
l) and a catalytic amount of 4- (dimethylamino) pyridine (DMA
P) was added. The resulting solution was gently warmed to 50 ° C. to complete dissolution. Two weeks later, the pyridine solution was put on ice, and the precipitated solid was collected by filtration. The obtained solid was dried over P ₂ O ₅ under reduced pressure, and 3 g (85%) of pale yellowish white powder was obtained.
I got

Example 67 3-Hydroxy-17β-[(3-pyridinylcarbonyl) oxy] -19-nor-17α-pregna-1,3,5 (1
Synthesis of 0) -trien-20-yne To 200 ml of a 0.5% solution of KHCO ₃ in methanol was added 2.0 g (3.9 mmol) of the product of Example 66. After 6 hours, the resulting slurry was diluted with 200 ml of water and the mixture was extracted with chloroform. The organic layers were combined, dried over MgSO _{4 and} concentrated under reduced pressure. The resulting oil was triturated with hexane to give 1.48 g (94%) of a white solid. NMR and UV
The structure of the title compound was confirmed by spectrum and elemental analysis.

Example 68 1-methyl-3-{[(19-nor-17α-pregna-1,
Synthesis of 3,5 (10) -trien-20-yn-17β-yl) oxy] carbonyl {pyridinium iodide 1.0 g (2.5 mmol) of the product of Example 67 was added to 50 ml of acetone, followed by methyl iodide. 2 ml was added. The reaction mixture was heated at reflux for 12 hours. When the precipitated solid is collected by filtration,
1.15 g (85%) of the quaternary salt represented by the structure below was obtained as a yellow solid.

The assigned structure was confirmed by UV and NMR spectra and elemental analysis.

Example 69 3-hydroxy-17β-{[(1-methyl-1,4-dihydropyridin-3-yl) carbonyl] oxy} -19-
Nor-17α-pregna-1,3,5 (10) -triene-20-
Synthesis of yne To a cooled suspension of 1.0 g (1.8 mmol) of the product of Example 68 in 100 ml of a mixed solvent of water and tert-butanol 50:50 was added NaH
0.73 g of CO ₃ and 0.96 g of Na ₂ S ₂ O ₄ were added. Reaction mixture
After stirring at 1 ° C for 1 hour, add 2
Extracted times. The combined organic extracts were dried over MgSO _{4 and} concentrated in vacuo to give 520 mg (6 mg) of the title compound as a yellow foam.
9%). The following structure was assigned to this product.

This structure was consistent with UV and NMR spectral values and elemental analysis.

Example 70 3,17β-bis [(3-pyridinylcarbonyl) oxy]
Synthesis of estra-1,3,5 (10) -triene 5.3 g of nicotinoyl chloride in 30 ml of dry pyridine at 0 ° C.
(0.03 mol), β-estradiol 2.0 g (0.0073mo
l) was added. After the reaction mixture was heated under reflux for 1 hour, it was poured onto 100 ml of ice water, and the deposited precipitate was collected by filtration.
The precipitate was dried under reduced pressure over P ₂ O _5, to give the title compound, mp 148~150 ℃ 3.18g (90%).

Example 71 1,1'-dimethyl-3,3 '-｛[(estradi-1,3,5 (1
Synthesis of 0) -triene-3,17β-diyl) dioxy] dicarbonyl dipyridinium iodide To 50 ml of acetone and 2 ml (0.032 mol) of methyl iodide, 2.0 g (0.004 mol) of the product of Example 70 was added. . The resulting solution was heated at reflux overnight. The deposited precipitate was separated by filtration, washed with acetone and dried, and represented by the following structure, having a melting point of 25.
2.75 g (88%) of the quaternary salt at 1-252 ° C. were obtained.

The above structure was confirmed by UV, NMR and elemental analysis.

Example 72 3,17β-bis {[(1-methyl-1,4-dihydropyridin-3-yl) carbonyl] oxy} estradi-1,3,5
Synthesis of (10) -triene 1 g (1.31 mmol) of the product of Example 71 was dissolved in 100 ml of dry acetonitrile. Nitrogen is bubbled through the solution,
(Phenylmethyl) -4- (aminocarbonyl) -1,2
0.28 g (1.31 mmol) of dihydropyridine were added and the reaction mixture was stirred at 0 ° C. for 1 hour. The solvent was reduced and the resulting solid was suspended in methylene chloride and filtered. The filtrate was chromatographed several times on a neutral alumina column prepared with methylene chloride. After purification, the solvent was removed under reduced pressure to give a solid foam. This product was represented by the following formula, and its structure was confirmed by UV, NMR and elemental analysis.

Example 73 3- (Phenylcarbonyloxy) -17β-[(3-pyridinylcarbonyl) oxy] -estradi-1,3,5 (1
Synthesis of 0) -triene 2.5 g (6.6 mmol) of estradiol benzoate was dissolved in 50 ml of dry pyridine, and then 1.66 g of nicotinic anhydride was dissolved.
And a catalytic amount of 4- (dimethylamino) pyridine (DMAP). The reaction mixture was stirred at room temperature for 5 days and then poured onto ice water. The deposited precipitate was collected by filtration and dried under reduced pressure.
3.01 g (94%) of the product as a white solid with a melting point of 151-154 ° C. were obtained.

Example 74 1-methyl-3-[{[(3-phenylcarbonyloxy) estra-1,3,5 (10) -trien-17β-yl]
Synthesis of [oxy @ carbonyl] pyridinium iodide 1.5 g (3.1 mmol) of the product of Example 73 was suspended in 2.5 ml of acetone. Then 2 ml of methyl iodide were added and the reaction mixture was heated at reflux overnight. The precipitated yellow solid (1.8 g,
93%) was collected by filtration and dried under reduced pressure. UV, NMR and elemental analysis confirmed that the product had the following assigned structure.

Example 75 3- (phenylcarbonyloxy) -17β-{[(1-
Synthesis of methyl-1,4-dihydropyridin-3-yl) carbonyl] oxydiestra-1,3,5 (10) -triene 1.2 g (1.93 mmol) of the quaternary salt prepared in Example 74
-Butyl alcohol / water was suspended in 100 ml of a 50:50 mixed solvent. Then, 0.81 g of NaHCO ₃ and 1.0 g of Na ₂ S ₂ O ₄ were added,
The reaction was allowed to continue for 1.5 hours. The resulting solution was extracted with methylene chloride, the organic phase was dried over MgSO _{4 and} concentrated under reduced pressure to give 650 mg of the title compound as a yellow foam. The structure of the product was confirmed by UV, NMR and elemental analysis. The following structure was assigned to this product.

Example 76 N- [2- {4- [bis (2-chloroethyl) amino]
Synthesis of butanoyloxydiethyl] -3-pyridinecarboxamide After dissolving 20 g (0.0657 mol) of chlorambutyl in 800 ml of dry acetonitrile, N- (2-hydroxyethyl)
13.1 g (0.079 mol) of -3-pyridinecarboxamide was added. Acetonitrile was added until the solution became clear.
The total amount of acetonitrile used at this stage was 850 ml. This solution was stirred while keeping the argon atmosphere, and 1.492 g of dicyclohexylcarbodiimide (0.
072 mol) and 4- (dimethylamino) pyridine (DMAP) 0.
802 g (0.0066 mol) were added. The reaction mixture was stirred overnight at room temperature under dry conditions, and the progress of the reaction was monitored by thin layer chromatography. After the elapse of the reaction time, the precipitated solid was filtered off and washed with 50 ml of cold acetonitrile. The filtrate was evaporated under reduced pressure at 30 ° C., and the obtained yellow solid was dissolved in a minimum amount (15 ml) of a mixed solvent of chloroform / tetrohydrofuran 8: 2 and charged into a column filled with 900 g of silica gel. The column was eluted with a mixture of chloroform / tetrahydrofuran 8: 2. The addition product and chlorambutyl eluted in the first 500 ml, then the desired ester was obtained by evaporation of the eluate under reduced pressure. The title compound was obtained in 82.7% yield as a yellow solid, mp 73-75 ° C. This was represented by the following structural formula.

Example 77 Synthesis of 1-methyl-3-[(N- {2- [4-({4-bis (2-chloroethyl) amino} phenyl) butanoyloxy] ethyl}) carbamoyl] pyridinium methanosulfenoate 2 g (0.04 mol) of the product of Example 76 were dissolved in 200 ml of dry acetonitrile. 0.613 g of dimethyl sulfate (0.
0486 mol) was added and the mixture was refluxed overnight. The reaction was followed by thin layer chromatography (chloroform / tetrahydrofuran 8: 2) until no unquaternized ester remained. The solvent was removed under reduced pressure, and the obtained residue was washed several times with dry ether. The remaining red viscous liquid was stored under an argon atmosphere until further use. 97.35% yield. The product was identified as the desired quaternary salt by NMR analysis. The following structural formula was assigned to this.

Example 78 1-methyl-3-[(N- {2- [4-({4- [bis (2-chloroethyl)] amino} phenyl) butanoyloxy] ethyl}) carbamoyl] -1,4-dihydropyridine 2.49 g (0.0043 mol) of the quaternary salt obtained in Example 77 was added to 350 m of water.
l. Nitrogen was bubbled through the solution throughout the reaction. After cooling this aqueous solution to 5 ° C. with a water bath, NaHCO ₃
2.17 g (0.026 mol) was added over 5 minutes, followed by the addition of 2.997 g (0.017 mol) of sodium dithionite over 10 minutes. The reaction mixture was kept at 5 ° C. for 120 minutes and then separated. The aqueous layer was extracted four times with ethyl acetate. The ethyl acetate extracts were combined, dried over MgSO ₄ and evaporated to dryness under reduced pressure. The resulting semi-solid was dried and washed several times with ether to give the title compound having the following structure, melting point 90-92 ° C
As a yellow solid.

Example 79 Synthesis of N- (4-hydroxycyclohexyl) -3-pyridinecarboxamide 5.05 g (0.033 mol) of trans-4-aminocyclohexanol hydrochloride was suspended in 50 ml of ethanol, and the reaction mixture was cooled to 10 ° C. While adding 1N NaOH33m to this suspension
l was added slowly. After the obtained homogeneous mixture was evaporated to dryness, the operation of adding a 50:50 mixed solvent of benzene and acetone and evaporating to dryness under reduced pressure was repeated three times. The dry solid thus treated was extracted with 100 ml of chloroform, filtered and the filtrate was evaporated to dryness. Trituration with residual ether and drying gave 3.50 g (91.23%) of free aminocyclohexanol, mp 111-112 ° C.

2.14 g of nicotinic acid (0.04 g in 75 ml of dry tetrohydrofuran)
17 mol) were suspended, and then 1.76 freshly distilled triethylamine was added. After cooling the resulting clear solution to −4 ° C. in an ice bath under an argon atmosphere, 1.88 g (0.014 mol) of ethyl chloroformate in 10 ml of tetrahydrofuran.
Was added at a rate such that the temperature did not exceed 0 ° C. After 2.0 g (0.017 mol) of the above free aminocyclohexanol was added to the cooled reaction mixture in powder form, the reaction mixture was heated to room temperature and stirred for 2 hours. The precipitated precipitate was separated by filtration, dissolved in hot water (28 ml), and recrystallized to give 3.35 g (85%) of the title compound having the following structure, melting point: 208-210 ° C
As fine colorless needle-like crystals.

The above structural example was confirmed by elemental analysis.

Example 80 N- {4- [4-(@ 4- [bis (2-chloroethyl)]
Synthesis of amino {phenyl) butanoyloxy] cyclohexyl} -3-pyridinecarboxamide 1.38 g (0.0045 mol) of chlorambutyl and N- (4-hydroxycyclohexyl) -3-pyridinecarboxamide
1.1 g (0.0049 mol) were mixed with 1.03 g (0.0459 mol) of dicyclohexylcarbodiimide and 55 mg (0.000453 mol) of 4- (dimethylamino) pyridine (DMAP) in 50 ml of freshly distilled acetonitrile. The reaction mixture was stirred for 2 days at room temperature in the presence of argon. The progress of the reaction was monitored by thin layer chromatography using an 8: 2 mixed solvent of chloroform / tetrahydrofuran. After the elapse of the reaction time, the solid thus separated was filtered off, and the filtrate was evaporated to dryness under reduced pressure at a low temperature. The residue was put on a silica gel column and eluted with chloroform / tetrahydrofuran 8: 2. The appropriate eluted fractions were combined and evaporated to dryness under reduced pressure. 1.86 g (81%) of a light creamy powdery product having the following structure and a melting point of 120-122 ° C. were obtained.

The structure of the product was confirmed by elemental analysis.

Example 81 Synthesis of 1-methyl-3- (N- {4- [4- (4-{[bis (2-chloroethyl)] amino} phenyl) butanoyloxy] cyclohexyl} carbamoyl) pyridinium methanosulfate 1 g (0.0019 mol) of the product of 80 was dissolved in 30 ml of dry acetonitrile and 0.249 g of dimethyl sulfate (0.
0019 mol) was added. The mixture was refluxed under an argon atmosphere until thin layer chromatography (silica column, chloroform / tetrahydrofuran 8: 2) indicated that the quaternization reaction was complete (about one and a half days). The solvent was removed under reduced pressure and the resulting orange residue was washed several times with anhydrous ether and evaporated under reduced pressure. 1.04 g of the desired quaternary salt (80.
6%) as a sticky yellow substance. This was represented by the following structural formula.

Example 82 Synthesis of 1-methyl-3- (N- {4- [4- (4-{[bis (2-chloroethyl)] amino} phenyl) butanoyloxy] cyclohexyl} carbamoyl) -1,4-dihydropyridine 0.34 g (0.0005 mol) of the quaternary salt obtained in Example 81 was dissolved in 0.5 ml of acetonitrile and absorbed in 20 ml of degassed water cooled to 0 ° C. (aerated with nitrogen gas during the reaction). .
Stir the resulting solution, add sodium bicarbonate 0.27 g
(0.003 mol) was added, followed by 0.37 (0.002 mol) of sodium dithionite and then 20 ml of ethyl acetate. After 90 minutes, the organic layer was separated and the aqueous layer
Extracted four times. The ethyl acetate extracts were combined, dried over sodium sulfate and evaporated under reduced pressure. The residual solid was washed several times with anhydrous ether and dried. The residue thus obtained was placed on a neutral alumina column and eluted with chloroform under pressure. Evaporation of chloroform from the eluate gave 0.18 g (65%) of the title compound represented by the following structural formula as a hygroscopic yellow solid.

The structure of the product was confirmed by UV analysis.

Example 83 Synthesis of N- (2-hydroxy) propyl-3-pyridinecarboxamide 4.29 g (0.039 ml) of nicotinic acid were suspended in 120 ml of dry tetrahydrofuran, and 4.04 g (0.039 mol) of freshly distilled triethylamine was once added thereto. Added. The resulting clear solution was cooled to -4 ° C in an ice bath under an argon atmosphere. Then, 4.33 g (0.039 mol) of ethyl chloroformate dissolved in 25 ml of tetrahydrofuran was added to the solution at 0 ° C.
Was added at a rate not to exceed. Next, 3 g (0.039 m) of 1-amino-2-propanol was added to the cooled reaction mixture.
ol) was added directly. The reaction mixture was warmed to room temperature and
Stirred for hours. The deposited precipitate was removed by filtration, and the filtrate was evaporated under reduced pressure. The resulting oily residue was washed several times with anhydrous ether and allowed to stand. 6.11 g of the title compound having the following structure (85
%) Was obtained as a hygroscopic, low melting white waxy solid with a melting point of 40 ° C.

Example 84 Synthesis of N- {2- [4-({4- [bis (2-chloroethyl)] amino} phenyl) butanoyloxy] propyl} -3-pyridinecarboxamide 1.0 g (0.003 mol) of chlorambutyl and N- 0.065 g of (2-hydroxy) propyl-3-pyridinecarboxamide
(0.0036 ml) in 0.68 of dicyclohexylcarbodiimide
g (0.003 mol) and 41 mg (0.003 mol) of 4- (dimethylamino) pyridine (DMAP) were mixed in 40 ml of freshly distilled acetonitrile. The reaction mixture was stirred for 2 days at room temperature in the presence of argon, and the progress of the reaction was monitored by thin-layer chromatography on a silica column using an 8: 2 mixture of methylene chloride / ethyl acetate. After the elapse of the reaction time, the precipitated solid was filtered, and the filtrate was evaporated to dryness under reduced pressure at 30 ° C. The residue was put on a silica gel column and eluted with methylene chloride / ethyl acetate 8: 2. The appropriate eluates were collected and combined and evaporated to dryness under reduced pressure. The title compound as a sticky substance was obtained in 84% yield (1.53 g). This was represented by the following structural formula.

Example 85 Synthesis of 1-methyl-3-[(N- {2- [4-({4- [bis (2-chloroethyl)] amino} phenyl) butanoyloxy] propyl}) carbamoyl] pyridinium methanosulfate 2.2 g (0.0047 mol) of the product of Example 84 are dissolved in 45 ml of dry acetonitrile and 0.59 g of dimethyl sulfate are dissolved.
(0.0047 mol) was added and the mixture was refluxed under an argon atmosphere. The progress of the reaction was followed by thin layer chromatography using methylene chloride / ethyl acetate 8: 2. After one and a half days, the solvent was removed by evaporation under reduced pressure, and the obtained orange residue was thoroughly washed with anhydrous ether and dried under reduced pressure. The desired quaternary salt product was obtained as a yellow sticky substance in a yield of 92.47%, which was represented by the following structural formula.

Example 86 1-methyl-3-[(N- {2- [4-({4- [bis (2-chloroethyl)] amino} phenyl) butanoyloxy] propyl}) carbomoyl] -1,4-dihydropyridine 2.39 g (0.004 mol) of the quaternary salt obtained in Example 85 was dissolved in 1 ml of acetonitrile, and 100 ml of degassed water (aerated with nitrogen gas during the reaction) cooled to 0 ° C. in an ice water bath. Was absorbed. While stirring the resulting solution, add sodium bicarbonate
2.03 g (0.024 mol) were added followed by 2.81 g (0.016 mol) of sodium dithionite. 60 ml of ethyl acetate was added to the resulting mixture. After the reaction was allowed to proceed for 90 minutes, the layers were separated and the aqueous layer was extracted three to four times with 30 ml portions of ethyl acetate. The ethyl acetate extracts were combined, dried over sodium sulfate and evaporated under reduced pressure. The residue was placed on a neutral aluminum column and eluted with chloroform under pressure. The appropriate eluate is collected and evaporated to yield a hygroscopic yellow solid.
% Yield. This product had the following structure:

The structure of the product was confirmed by UV analysis.

Example 87 Synthesis of N- (2-hydroxy-2-phenyl) ethyl-3-pyridinecarboxamide 1.79 g (0.014 mol) of nicotinic acid was suspended in 60 ml of dry tetrahydrofuran, and 1.48 g (0.014 mol) of freshly distilled triethylamine was added thereto. mol) was added. The resulting clear solution was cooled to −4 ° C. in an ice bath and argon was continuously bubbled through it. Then, 1.58 g (0.014 mol) of ethyl chloroformate dissolved in 10 ml of tetrahydrofuran was added at such a rate that the temperature of the solution did not exceed 0 ° C. Next, 2-
2.0 g (0.014 mol) of amino-1-phenylethanol were added as a solution in 5 ml of tetrahydrofuran. The reaction mixture was warmed to room temperature and stirred for 2 hours. The deposited precipitate was removed by filtration, and the filtrate was evaporated under reduced pressure to have a melting point of 12 having the following structure.
3.22 g (91.1%) of a white crystalline solid at 2-124 ° C. was obtained.

Elemental analysis confirmed the structure of the product.

Example 88 N-(｛2-phenyl-2- [4-(｛4- [bis (2
Synthesis of -chloroethyl)] amino {phenyl) butanoyloxy] {ethyl) -3-pyridinecarboxamide 1.0 g (0.003 mol) of chlorambutyl and 0.88 g of N- (2-hydroxy-2-phenyl) ethyl-3-pyridinecarboxamide ( 0.003 mol) were mixed with 0.38 g (0.003 mol) of dicyclohexylcarbodiimide and 41 mg (0.0003 mol) of 4- (dimethylamino) pyridine (DMAP) in 35 ml of freshly distilled acetonitrile. The reaction mixture was stirred at room temperature for 3 days in the presence of argon. The success of the reaction was followed by thin-layer chromatography using an 8: 2 mixed solvent of methylene chloride / ethyl acetate. After the reaction, the precipitated solid was removed by filtration, and acetonitrile was evaporated under reduced pressure. The obtained residue was put in a silica gel column and eluted with methylene chloride / ethyl acetate 8: 2. The appropriate eluates were collected, combined and evaporated under reduced pressure to give 1.21 g (70%) of a pale tan powder of melting point 99-101 ° C. and having the structure shown below.

Example 89 1-methyl-3-[(N- {2-phenyl-2- [4-
Synthesis of ({4- [bis (2-chloroethyl)] amino} phenyl) butanoyloxy] {ethyl) carbamoyl] pyridinium methanosulfate 0.5 g (0.00094 mol) of the product of Example 88 was dissolved in 20 ml of dry acetonitrile. , Dimethyl sulfate 0.12g
(0.00094 mol) was added. After the mixture was refluxed under an argon atmosphere for 2 days, the solvent was removed by evaporation under reduced pressure. The resulting residue was washed several times with anhydrous ether, dried, and 0.54 g of a sticky yellow product represented by the following structural formula.
(91%).

Example 90 1-methyl-3-[(N- {2-phenyl-2- [4-
({4- [bis (2-chloroethyl)] amino} phenyl) butanoyloxy] {ethyl) carbamoyl] -1,
Synthesis of 4-dihydropyridine 0.53 g (0.0008 mol) of the quaternary salt obtained in Example 89 was added in 0.5 ml.
Of acetonitrile and absorbed in 20 ml of degassed deionized water cooled to 0 ° C. While stirring the resulting solution at 0 ° C., sodium bicarbonate was added, followed by 0.56 g (0.0032 mol) of sodium dithionite. Further 20 ml of ethyl acetate was added and the reaction was allowed to proceed for 2 hours.
The organic layer was separated and the aqueous layer was extracted several times with ethyl acetate until no color was observed in the organic layer (total volume 70 ml). The ethyl acetate extracts were combined, dried over sodium acetate,
Evaporated under reduced pressure. The residue was put on a neutral alumina column and eluted with chloroform. The appropriate eluate was collected and the chloroform was evaporated, yielding 0.2 g (45%) of a hygroscopic orange-yellow compound of the following formula.

The structure of the product was confirmed by UV spectral analysis.

Example 91 Synthesis of N- (2-hydroxyethyl) -3-pyridinecarboxamide A solvent-free mixture of 6.1 g (0.10 mol) of 2-aminoethanol and 15.1 g (0.10 mol) of ethyl nicotinate was refluxed overnight. . Upon cooling the mixture to room temperature, the product precipitated as a crystalline solid. This was filtered off, washed with ether and then recrystallized from 2-propanol / ether. The final product was collected by vacuum filtration and washed with ether. The weight of the obtained white compound after drying was 10.7.
g, yield 64.5%, melting point 88.5-89.5 ° C (literature value: 92
° C).

Example 92 Synthesis of (+) N- [2- (6-methoxy-α-methyl-2-naphthalenylacetoxy) ethyl] -3-pyridinecarboxamide 2.30 g (10.0 mmol) of naproxen was added to acetonitrile 1
2.30 g of dicyclohexylcarbodiimide (11.0 m
mol) and 122 mg of 4- (dimethylamino) pyridine (1.
Using 1.7 mmol of the product of Example 91 (10.0 mmol)
l). The reaction mixture was stirred at room temperature for 48 hours. The deposited precipitate was filtered off, rinsed with acetonitrile,
Upon drying, it weighed 2.3 g. The solvent was removed from the solution under reduced pressure and the remaining clear oil was stirred with anhydrous ether. The resulting white solid was filtered under reduced pressure, washed with ether, and air-dried. The amount of the crude product was 2.80 g. This compound was recrystallized from 2-propanol. The final product was filtered off and washed with 0.5% aqueous sodium bicarbonate, water, and finally with ether. This compound was dried in a desiccator over P ₂ O ₅ . The amount of product after recrystallization is 2.
The amount was 40 g, the overall yield was 63.4%, and the melting point was 79 to 82 ° C.

Example 93 Synthesis of N- (2-{[1- (p-chlorobenzoyl) -5-methoxy-2-methyl-3-indolyl] acetoxy} ethyl) -3-pyridinecarboxamide 1.79 g (5.00 mmol) of indomethacin Reaction with 0.820 g (5.00 mmol) of the product of Example 91 was carried out using 1.10 g (5.50 mmol) of dicyclohexylcarbodiimide as binder and acetonitrile as solvent. The first two reaction components were completely dissolved in the solvent and the resulting solution was
Cooled to ° C. Dicyclohexylcarbodiimide was added and the mixture was stirred overnight. The reaction was allowed to continue for 48 hours.
The precipitated precipitate (1.2 g) was removed by filtration under reduced pressure. Removal of the solvent from the filtrate under reduced pressure gave an oily residue. This product was solidified by stirring with anhydrous ether. The precipitate was filtered, air dried and recrystallized from ethanol / ether. The final product was filtered under reduced pressure, washed with ether and air-dried. The yield of the product was 1.65 g, yield 65.2%, melting point 123-125 ° C.

Example 94 Synthesis of 1-methyl-3- {N- [2- (6-methoxy-α-methyl-2-naphthalenylacetoxy) ethyl] carbamoyl} pyridinium iodide 1.0 g of naproxen ester prepared in Example 92 (2.6m
mol) of methyl iodide 2.3 in 45 ml of acetone.
g (16 mmol). The resulting solution was heated at reflux for 20 hours. 1.1g of methyl iodide (8.0m
mol). After the reaction was further continued for 4 hours, the precipitated product was separated by filtration. The obtained pale yellowish white powder was dried. The yield of this product was 2.2 g, which was found to be analytically pure and did not require recrystallization. The solvent was removed from the acetone solution of the filtrate, and the residue was solidified with anhydrous ether. The obtained deep yellow powder was dissolved in water, and ether (4
× 30 ml). Then, the water was removed under reduced pressure to obtain 0.2 g of a powder with a pale yellow coloration. The overall yield of the reaction was 93%, mp 169-170 ° C. The product was represented by the following structural formula, and its structure was confirmed by UV, MNR and elemental analysis.

Example 95 Synthesis of 1-methyl-3- [N- (2-{[1- (p-chlorobenzoyl) -5-methoxy-2-methyl-3-indolyl] acetoxy} ethyl) carbomoyl] pyridinium iodide 0.50 g of indomethacin ester obtained in Example 93 (1.0 mm
ol) in 1.7 g of methyl iodide (12 g) in acetone.
mmol). The reaction was heated at reflux overnight.
Removal of the solvent under reduced pressure gave a yellow solid. The product was recrystallized using ethanol and only a small amount of ether. Small, pale yellow, moldy crystals were obtained. The yield of the reaction product was 043 g, and the yield of the purified product was 66%. Melting point 178-179 [deg.] C. By UV, MNR and elemental analysis, it was confirmed that the product was represented by the following structural formula.

Example 96 Synthesis of 1-methyl-3- {N- [2- (6-methoxy-α-methyl-2-naphthalenylacetoxy) ethyl] carbamoyl} -1,4-dihydropyridine Prepared in Example 94 780 mg (1.5 mmol) of the quaternary salt were dissolved in a mixed solvent of 200 ml of degassed deionized water and 10 ml of acetonitrile. Sodium dithionite 780mg (4.5mm
ol) and 630 mg (7.5 mmol) of sodium bicarbonate,
The solution was added at room temperature. The reaction was continued for 1 hour while slowly passing nitrogen gas through the solution. The incompletely precipitated product was repeatedly extracted with ether (8 × 30 ml). The extracts were combined, washed with water (25 ml) and dried over magnesium sulfate. The drying agent was removed by filtration, and the solvent was removed from the filtrate under reduced pressure. The operation of dissolving the obtained oily residue in 5 ml of methylene chloride and removing the solvent under reduced pressure was repeated three times. The obtained foam was washed with anhydrous ether (3 ml), and the solvent was removed under reduced pressure. The amount of the final product was 390 mg and the yield was 66%. This hygroscopic solid foam was stored at -100 ° C under a nitrogen atmosphere. Its structural formula is as follows: UV, NM
The structure was confirmed by R and elemental analysis.

Example 97 1-methyl-3- [N- (2-{[1- (p-chlorobenzoyl) -5-methoxy-2-methyl-3-indolyl] acetoxy} ethyl) carbamoyl] -1,4-dihydropyridine Synthesis of the indomethacin quaternary salt 140 mg prepared in Example 95
(0.22 mmol) in a minimum amount of water: acetonitrile (8: 2)
Was dissolved. The water was flushed with nitrogen for 20 minutes before use. While stirring this solution at 0 ° C., 91 mg (1.1 mmol) of sodium bicarbonate and 110 mg of sodium dithionite were added.
mg (0.65 mmol) was added. The solution was then warmed to room temperature. The reaction was continued for about one hour. Some of the product precipitated during the reaction. This was dissolved in ethyl ether. The aqueous layer was extracted several times with ether until no yellow color transferred to the organic layer. The ether layers were combined, dried over magnesium sulfate, filtered, and the ether was removed in vacuo. The operation of dissolving the remaining oil in 10 ml of acetone and removing the solvent under reduced pressure was repeated twice to obtain a dry foam. The amount of this final product was 92 mg. Yield 82%, mp 60-65 ° C. The structural formula of the product is as follows, and the structure was confirmed by UV, NMR and elemental analysis.

Example 98 N-[(1-hydroxycyclohexyl) methyl] -3
Synthesis of pyridinecarboxamide 1.48 g (0.012 mol) of nicotinic acid were suspended in 50 ml of dry tetrahydrofuran, and 2.44 g (0.014 mol) of freshly distilled triethylamine were added thereto with stirring. The resulting clear solution was cooled to -4 ° C in an ice bath under an argon atmosphere.
And cooled. Then 1.3 g (0.012 mol) of ethyl chloroformate dissolved in 10 ml of tetrahydrofuran were added at such a rate that the temperature of the reaction mixture did not exceed 0 ° C. Next, 2.0 g (0.012 mol) of 1-aminomethyl-1-cyclohexanol hydrochloride was directly added to the cooled reaction mixture as a powder. After the reaction mixture was warmed to room temperature and stirred for 2 hours, the precipitated triethylamine hydrochloride was removed by filtration, and the filtrate was evaporated under reduced pressure to obtain a white solid. This solid was recrystallized from water, washed with acetone and ether and dried.
85% yield (2.4g) of the title compound melting at around 110 ° C
I got it. This was confirmed by elemental analysis to be a compound represented by the following structural formula.

Example 99 Synthesis of N-({1- [4-({4- [bis (2-chloroethyl) amino} phenyl) butanoyloxy] cyclohexyl} methyl) -3-pyridinecarboxamide 1.18 g (0.0038 mol) of chlorambutyl 0.99 g (0.004 mol) of N-[(1-hydroxycyclohexyl) methyl] -3-pyridinecarboxamide, along with 0.8 g (0.0038 mol) of dicyclohexylcarbodiimide and 47 mg (0.00038 mol) of 4- (dimethylamino) pyridine (DMAP), Mix in 60 ml of freshly distilled acetonitrile. The reaction mixture was stirred at room temperature under an atmosphere of argon for 7 days. After the reaction time had elapsed, the precipitated solid was filtered off, and the filtrate was evaporated to dryness under reduced pressure at a low temperature. The residue was placed on a silica gel column and eluted first with methylene chloride / ethyl acetate 8: 2 and then with chloroform / tetrahydrofuran 8: 2. The appropriate eluates were collected and combined and evaporated to dryness under reduced pressure. The title compound was obtained as a pale yellow solid with a melting point of 92-94 ° C. in 26% yield. This is represented by the following structural formula,
Elemental analysis confirmed the structure.

Example 100 1-Methyl-3- [N-({1- [4- (4-{[bis (2-chloroethyl)] amino} phenyl) butanoyloxy] cyclohexyl} methyl) carbamoyl] pyridinium methanosulfate Synthesis The product of Example 99 dissolved in 25 ml of dry acetonitrile
0.69g (0.0013mol), 0.17g of dimethyl sulfate
(0.0013 mol) was added. The mixture was heated at reflux until the reaction was complete (about 2 days) as determined by thin layer chromatography using chloroform / tetrahydrofuran 8: 2. The solvent was evaporated off under reduced pressure and the orange residue obtained was washed several times with anhydrous ether and dried. The product obtained is 0.72 g (85%) of a sticky yellow substance which is a compound of the structure

Example 101 1-methyl-3- [N-({1- [4- (4-{[bis (2-chloroethyl)] amino} phenyl) butanoyloxy] cyclohexyl} methyl) carbamoyl] -1,
Synthesis of 4-dihydropyridine 0.5 ml of 0.78 g (0.0012 mol) of the quaternary salt obtained in Example 100
In acetonitrile, cooled to 0 ° C., and taken up in 20 ml of water which had been degassed by bubbling nitrogen through. While stirring the resulting solution, sodium bicarbonate 0.61 g (0.0
072 mol) and then 0.84 g of sodium dithionite
(0.0048 mol) and 20 ml of ethyl acetate were added. After allowing the reaction to proceed for 75 minutes, the layers were separated and the aqueous layer was ethyl acetate 20 ml.
Each extraction was performed three to four times. The organic extracts were combined, dried over sodium sulfate and evaporated under reduced pressure. The residue was placed on a neutral alumina column and eluted with chloroform under pressure. The appropriate eluate was collected and evaporated to give the product as a sticky yellow material (0.31 g, 49%). This product had the following structure:

Example 102 Synthesis of 1-methyl-3-{[2- (9-guanylmethoxy) ethoxy] carbonyl} pyridinium iodide
Et al., Synthetic Communications , 17 (4) , 451-455 (1
987).

Acyclovir 1.25 ml of freshly distilled dry pyridine 1.
0 g (4.4 mmol) was dissolved in anhydrous trigonelline.
2.27 g (4.4 mmol) of diiodide and the amount of catalyst (5.4 mg, 4 mmol)
Of 4- (dimethylamino) pyridine (DMAP). The resulting suspension was stirred at room temperature under argon atmosphere for 4 days. As the reaction proceeded, the orange color of the acid anhydride turned yellow. After all the acyclovir has been consumed, the reaction is stopped, the precipitate (containing trigonelline by-produced in addition to the ester of the target product) is filtered off and washed with acetone and ether to remove DMAP. did. Then
The yellow solid was stirred in dry methanol at room temperature to remove trigonelline, unreacted anhydride and acyclovir. The title compound was obtained in 87% yield (1.82 g). 201-202 ° C. NMR and UV analysis confirmed that the product had the structure shown below.

Example 103 Synthesis of 1-methyl-3-{[2- (9-guanylmethoxy) ethoxy] carbonyl} -1,4 dihydropyridine 1.58 g (3.3 mmol) of the product of Example 102 was degassed in water 120
To the solution dissolved in ml, 1.69 g (20.1 mmol) of NaHCO ₃ was added in one portion. While the mixture was stirred at 0 ° C., 2.33 g (13.18 mmol) of sodium dithionite was added over 5 minutes. During the reaction, nitrogen gas was kept flowing through the flask.
The resulting dihydropyridine product is insoluble in water,
Cream crystals formed on the aqueous layer. The crystals were filtered off and washed first with ice-cold water and then with anhydrous ether. −
And dried by P ₂ O ₅ in a desiccator held at 15 ° C., the title compound, mp 163~165 ℃ 0.626g (54%) was obtained. NMR and UV analysis confirmed that the product had the structure shown below.

Example 104 Synthesis of 5'-pivaloyl trifluorothymidine A solution of 150 mg of trifluorothymidine in 5 ml of pyridine was stirred, and a solution of 90 mg of piperoyl chloride in 1 ml of pyridine was added thereto while cooling. After stirring at room temperature for 10 hours, the reaction mixture was poured into 20 ml of ice water and extracted with 50 ml of ethyl acetate. The extract was washed with water and dried over sodium sulfate. Ethyl acetate was removed, and the residue was purified by silica gel column chromatography using chloroform / methanol 20: 1 as an eluent. After recrystallization from a mixed solvent of ether and n-hexane, a melting point of 13
The title compound was obtained at 0-132 ° C.

Example 105 Synthesis of 3 '-(3-pyridylcarbonyl) -5'-pivaloyltrifluorothymidine 1.0 g of nicotinoyl hydrochloride was added. After stirring the reaction mixture at room temperature for 3 days, it was poured into 100 ml of ice water and extracted with 100 ml of ethyl acetate. The extract was washed with water, dried over anhydrous sodium sulfate and evaporated under reduced pressure to give an oil. n-
Crystallization from hexane gave 500 mg (87%) of colorless needles mp 175-177 ° C. The structure of this product is shown by the following formula, and the structure was further confirmed by NMR spectrum analysis.

Example 106 3 '-(1-Methyl-3-pyridiniumcarbonyl)-
Synthesis of 5'-pivaloyltrifluorothymidine iodide 1.0 g of methyl iodide was added to 440 mg of the product of Example 105 dissolved in 10 ml of acetone. After the mixture was heated under reflux for 10 hours, the precipitated precipitate was collected by suction filtration.
550 mg of the expected product are obtained as yellow leaf-like crystals, mp 188-190 ° C. (decomposition). NMR analysis confirmed that the product had the structure shown below.

Example 107 Synthesis of 3 '-(1,4-dihydro-1-methyl-3-pyridinylcarbonyl) -5'-pivaloyltrifluorothymidine Example 106 was mixed with a mixed solvent of 20 ml of water and 20 ml of ethyl acetate. Stir the solution in which 100 mg of the product was dissolved, and add NaHCO ₃ 64m
g and 115 mg of Na ₂ S ₂ O ₄ were added under a stream of nitrogen gas. After stirring the obtained mixture at room temperature for 1 hour, the organic layer was separated and washed with water. The organic layer was dried over anhydrous Na ₂ SO _{4 and} evaporated under reduced pressure. The residue was triturated with a mixture of ether and n-hexane, and the precipitated yellow needle crystals were collected by suction filtration (50 mg, 62%). The melting point of this product was 168-170 ° C. NMR analysis confirmed that the product had the structure shown below.

Example 108 Synthesis of 3′-azido-3′-deoxy-5 ′-(3-pyridylcarbonyl) thymidine 1.18 g (4.42 mmol) of azidothymidine, nicotinic anhydride
A mixture of 1.11 g (4.86 mmol) and 0.15 g (1.22 mmol) of N- (dimethylamino) pyridine was added to 50 ml of pyridine.
And mixed. The reaction mixture was stirred overnight at room temperature. The resulting colorless and transparent reaction mixture was concentrated under reduced pressure to an opaque semi-solid state, and triturated with ether overnight. The resulting suspension was filtered and dried to give 1.97g of solid. Next, 1.0 g of the solid was subjected to chromatography on 20 g of silica gel using 10% ethanol / chloroform as an eluent. The desired fraction was separated to obtain 0.53 g of a white foam. This was crystallized from a mixed solvent consisting of ethanol, diethyl ether and hexane. 138.5-14
A product at 1.5 ° C. was obtained, which was confirmed by NMR and IR to be of the structure shown below.

Example 109 Synthesis of 3'-azido-3'-deoxy-5 '-[(1-methyl-3-pyridinium) carbonyl) thymidine iodide 0.53 g (2.0 mmol) of azidothymidine, 1.02 g (2.2 mmol of anhydrous trigonelline diiodide) ) And 67 mg (0.5 mmol) of N- (dimethylamino) pyridine were mixed in 25 ml of pyridine. The reaction mixture was stirred at room temperature for 5 days and then filtered. The filtrate was concentrated under reduced pressure until a residue was obtained, and the residue was triturated with acetone overnight. The resulting suspension was filtered, and the filtrate was concentrated under reduced pressure to obtain a foam. The foam was treated with water and then filtered to remove a small amount of insolubles. The filtrate was concentrated under reduced pressure to give a glassy yellow solid (0.50 g, 49%)
I got NMR and UV analysis confirmed that the product had the structure represented by the following formula.

Example 110 Synthesis of 3'-azido-3'-deoxy-5 '-[(1-methyl-1,4-dihydro-3-pyridinyl) carbonyl] thymidine Crude azidothymidine prepared by the method described in Example 109 1.45 g (2.82 mmol) of the quaternary derivative was dissolved in 50 ml of water and filtered. The filtrate was cooled in an ice bath and saturated with argon. Then 100 ml of ethyl acetate and 2.90 g of NaHCO ₃
Was added, followed 5 minutes later by 1.45 g of Na ₂ S ₂ O ₄ .
After allowing the reaction to proceed for 1 hour, the ethyl acetate layer was separated, and another ethyl acetate was added to continue the reaction. This operation was repeated to obtain three types of organic extracts with a total reaction time of 3 hours. These extracts were combined, concentrated under reduced pressure to give 1.01 g (9
2%) of foam. This foam was crystallized from methanol to obtain the title compound having the following formula and a melting point of 138 to 140 ° C. Its structure was confirmed by elemental analysis and NMR and UV.

Example 111 Synthesis of dopamine dipivalate oxalate To a stirred mixture of 28.1 g of pivaloyl chloride and 150 ml of trifluoroacetic acid was added 18.01 g of dopamine hydrobromide. After stirring this mixture for 2 hours, 14 ml of water was added and the mixture was concentrated under reduced pressure. The remaining oil was dissolved in chloroform and washed with cold 10% aqueous KHCO ₃ until the evolution of CO ₂ ceased. The layers were separated, the chloroform layer was washed with water, dried over MgSO ₄ , filtered and evaporated to dryness. The residue was ethyl acetate 10
It was dissolved in 0 ml and 7 g of oxalic acid was added together with 100 ml of ethyl acetate. The resulting solution was filtered to remove insolubles and 1.6 g of oxalic acid in 25 ml of ethyl acetate was added. The mixture was concentrated under reduced pressure and cooled. The precipitated crystals were separated by filtration to obtain 13 g of the title compound. 5.9 g of crystals due to cooling of mother liquor
I got This product was represented by the following formula.

Example 112 N- {β- [3,4-bis (pivalyloxy) phenyl]
Synthesis of chloromethyl ethyl diaminocarboxylate 822 mg (2 mmol) of dopamine dipivalate oxalate was suspended in 15 ml of dry tetrahydrofuran. 289 ml (1 mmol) of triethylamine were added and the mixture was stirred for 15 minutes, after which a further 278 ml (1 mmol) of triethylamine was added. Upon addition of 390 mg (6 mmol) of ClCO ₂ CH ₂ Cl, a heavy white precipitate immediately precipitated and gas was evolved. After stirring the reaction mixture at room temperature overnight, the precipitate was filtered off and the filtrate was filtered with 10 ml of 0.
Washed with 1M hydrochloric acid. The organic layer was then dried over magnesium sulfate and evaporated to dryness, yielding 1.1 g of a golden oil represented by the following structural formula.

The structure of this product was confirmed by elemental analysis.

Example 113 N- {β- [3,4-bis (pivalyloxy) phenyl]
Synthesis of ethyl @ aminocarbonyloxymethyl-3-pyridinecarboxylate 1.26 g (.04 mmol) of the chloromethyl carbamate derivative obtained in Example 112 was mixed with 10 ml of dry dimethylformamide, and the mixture was treated with 375 mg (3.04 mmol) of nicotinic acid. )
And 445 ml of triethylamine (5% excess) at room temperature for 15 ml
Was added to a solution obtained by premixing in dry dimethylformamide. After stirring the reaction mixture for 4 days, the deposited precipitate was filtered off. The filtrate was evaporated to dryness and the residue was dissolved in 20 ml of methylene chloride. The solution was washed twice with 10 ml portions of water. The solvent was removed under reduced pressure to obtain the title compound represented by the following formula.

The structure of this compound was confirmed by NMR.

Example 114 N- {β- [3,4-bis (pivalyloxy) phenyl]
Ethyl @ aminocarbonyloxymethyl-1-methyl-
Synthesis of 3-pyridinium carboxylate iodide 860 mg (1.78 mmol) of the product of Example 113 are mixed with 15 ml of dry acetonitrile and the mixture is treated with 223 ml of methyl iodide.
(3.56 mmol). The resulting mixture was stirred at room temperature for 6 hours, a further 223 ml (3.56 mmol) of methyl iodide was added and the mixture was stirred overnight. Then, evaporation to dryness afforded the title compound of the following formula as an orange-red oil.

The structure of this product was confirmed by NMR analysis.

Example 115 N- {β- [3,4-bis (pivalyloxy) phenyl]
Synthesis of ethyl @ aminocarbonyloxymethyl-1,4-dihydro-1-methyl-3-pyridinecarboxylate 54 mg of the quaternary salt prepared in Example 114 in 10 ml of water (0.0 mg
84 mmol) were treated with 30 mg (4 eq) of NaHCO ₃ , 60 mg (4 eq) of Na ₂ S ₂ O ₄ and 20 ml of ethyl acetate at 0 ° C. under a nitrogen atmosphere. After allowing the reaction to proceed for 1 hour and 20 minutes, the aqueous layer and the organic layer were separated, and the aqueous layer was extracted again with 20 ml of ethyl acetate. The organic layers were combined and dried over magnesium sulfate. The solvent was removed under reduced pressure and the resulting red-orange oil was dissolved in chloroform and incompletely purified from a short neutral alumina column by elution with chloroform. The desired fractions were subjected to silica thin layer chromatography on silica using chloroform / acetone 80:20. The highest band was recovered as the title compound represented by the structure below.

The identity of this product was confirmed by measuring its ability to release dopamine from plasma and brain homogenate by HPLC (high pressure liquid chromatography).

Example 116 Synthesis of 9-fluoro-11β, 17-dihydroxy-16α-methyl-21-[(3-pyridinylcarbonyl) oxy] pregna-1,4-diene-3,20-dione Dexamethasone 1 g (2.5 mmol ) Was dissolved in 50 ml of dry pyridine. Add 680 mg of nicotinic anhydride (3.0 mm
ol) and traces of 4- (dimethylamino) pyridine (DMA
P) and added. After allowing the reaction to proceed for 4 hours, the reaction mixture was poured into ice water and refrigerated overnight. The precipitated solid was separated by filtration and dried to obtain 1.08 g (87%) of a product represented by the following structural formula and having a melting point of 262 to 265 ° C.

The structure was confirmed by elemental analysis.

Example 117 1-methyl-3-{[(9-fluoro-11β, 17-dihydroxy-16α-methylpregna-1,4-diene-3,20
Synthesis of -dione-21-yl) oxy] carbonyl {pyridinium iodide 0.74 g (1.5 mmol) of the product of Example 116 was dissolved in 50 ml of acetone, and 2 ml of methyl iodide was added thereto. Then, to increase the solubility, a small amount (10 ml) of CH ₃ NO ₃
Was added. After allowing the reaction to proceed for 2 days, the precipitated solid was separated by filtration to obtain 0.54 g (yield: 56%) of the title compound represented by the following formula and having a melting point of 218 to 221 ° C.

The structure of the product was confirmed by elemental analysis.

Example 118 9-fluoro-11β, 17-dihydroxy-16α-methyl-21-｛[(1-methyl-1,4-dihydropyridine-3
-Yl) carbonyl] oxydpregna-1,4-diene-3,20-dione The steroidal system prepared in Example 117 according to the general reduction method described in Example 11 of U.S. Pat. No. 4,617,298. Using 0.78 mmol of the quaternary salt, 0.33 g of NaHCO ₃ and 0.41 g of Na ₂ S ₂ O ₄ , the reaction was carried out in a 50% aqueous methanol solution at 0 ° C. while purging with nitrogen. A product represented by the following structural formula was obtained.

Example 119 Synthesis of 11β, 17-dihydroxy-21-[(3-pyridinylcarbonyl) oxy] pregn-4-en-3,20-dione 2 g (5.5 mmol) of hydrocortisone were dissolved in 50 ml of dry pyridine. 1.38 g of nicotinic anhydride (6.0
5 mmol) and traces of 4- (dimethylamino) pyridine (D
MAP) was added and the reaction was allowed to proceed at room temperature for 4 hours. The pyridine solution was poured into ice water, and the precipitated solid was separated by filtration. The solid was dried under reduced pressure over P ₂ O ₅ to obtain 2.4 g (93%) of the title compound represented by the following structural formula.

This structure was confirmed by elemental analysis and UV spectrum analysis.

Example 120 Synthesis of 1-methyl-3-{[(11β, 17-dihydroxypregn-4-en-3,20-dione-21-yl) oxy] carbonyl} pyridinium iodide 1 g of product of Example 119 (2.1 mmol) was dissolved in 50 ml of acetone, and 4 ml of mesyl iodide was added thereto. The solution was stirred overnight at reflux temperature. Removal of the solvent gave the title compound as a yellow powder in 98% yield. Elemental analysis confirmed that the product had the structure shown below.

Example 121 11β, 17-dihydroxy-21-｛[(1-methyl-1,4-
Synthesis of dihydropyridin-3-yl) carbonyl] oxydpregn-4-en-3,20-dione Steroids prepared in Example 120 according to the general reduction method described in Example 11 of U.S. Patent No. 4,617,298. Using 0.8 mmol of quaternary, 0.34 g of NaHCO ₃ and 0.42 g of Na ₂ S ₂ O ₄ , the reaction was carried out in a 50% aqueous methanol solution at 0 ° C. while purging with nitrogen. A product represented by the following structural formula was obtained.

Example 122 Synthesis of N- (2-chloroethyl) -N '-[2- (3-pyridinecarbonyloxy) ethyl] -N-nitrosourea 1.25 g (52 mmol) of nicotinic acid 2-aminoethyl ester diacid acid A solution of 2 g (6 mmol) of 2,4,5-trichlorophenyl-N- (2-chloroethyl) -N-nitrosocarbamate in 40 ml of pyridine was stirred at room temperature under a nitrogen atmosphere for 24 hours. The reaction was performed by thin layer chromatography (silica, chloroform / ethyl acetate 1: 1, Rf0.26)
Was monitored by. After completion of the reaction, the solvent was removed under reduced pressure, and the residue was subjected to chromatography using a silica gel column. First, elution was performed with benzene to remove unreacted nitrosocarbamate compounds and by-product trichlorophenol, and then with chloroform. To give the desired product. The resulting oil was solidified in a freezer. This product had a melting point of 63-64 ° C. and was represented by the following structural formula.

Example 123 N- (2-chloroethyl) -N '-[2- (1-methyl-3-pyridiniumcarbonyloxy) ethyl] -N-
Synthesis of Nitrosourea Iodide A solution of 1.5 g (5 mmol) of the product of Example 122 in 40 ml of tetrahydrofuran was treated with excess methyl iodide. The mixture was stirred at 50 ° C. for 4 hours. The fine crystalline yellow solid thus obtained (1.8 g, 82%) has a melting point of 12%.
The temperature was 0 to 121 ° C., and it was confirmed by elemental analysis that the structure was represented by the following structure.

Example 124 Synthesis of N- (2-chloroethyl) -N '-[2- (1,4-dihydro-1-methyl-3-pyridinecarbonyloxy) ethyl] -N-nitrosourea Prepared in Example 123 1.48 g (1.1 mmol) of a quaternary salt type nitrosourea derivative and 0.23 g (1 mmol) of 1-benzyl-1,2-dihydroisonicotinamide were added to anhydrous methanol 25.
The solution dissolved in ml was stirred at 0 ° C. under a nitrogen atmosphere for 4 hours. The precipitated solid was separated by filtration and washed with methanol and ether. This solid was analyzed by NMR for 1-benzyl-4-
It was identified as carbamoylpyridinium iodide. The filtrate was evaporated under reduced pressure at about 30 ° C., and the residue was suspended in methylene chloride. The precipitated solid was filtered and washed with methylene chloride. The filtrate was evaporated under reduced pressure and the residue was dissolved in chloroform. Flash chromatography was performed on a short column of neutral alumina using chloroform as eluent, and the chloroform solution containing the desired product was evaporated under reduced pressure to give 0.2 g (63%) of a gummy residue. This is NM
By R, the product was identified as the target product represented by the following formula.

The compound thus obtained easily reduced the alcohol solution of silver nitrate.

Example 125 Synthesis of N- (2-fluoroethyl) -N '-[2- (3-pyridinecarbonyloxy) ethyl] -N-nitrosourea 1.5 g of nicotinic acid 2-aminoethyl ester dihydrochloride
(6.3 mmol) and 2,4,5-trichlorophenyl-N- (2-
Fluoroethyl) -N-nitrosocarbamate 2.18 g
(6.9 mmol) in 50 ml of pyridine was stirred at room temperature under a nitrogen atmosphere for 24 hours. The reaction was analyzed by thin layer chromatography (silica, chloroform / ethyl acetate 1: 1, R
f0.25). After completion of the reaction, the solvent was removed under reduced pressure, and the residue was subjected to chromatography using a silica gel column. First, elution was performed with benzene to remove unreacted nitrosocarbamate compounds and by-product trichlorophenol, and then with chloroform. To elute the desired product. 1.56 g (87.4%) of the title compound having a melting point of 75-77 ° C. were obtained, which was represented by the following structural formula.

Example 126 N- (2-fluoroethyl) -N '-[2- (1-methyl-2-pyridiniumcarbonyloxy) ethyl] -N
Synthesis of nitrosourea iodide A solution of 1.56 g (5.4 mmol) of the product of Example 125 in 40 ml of tetrahydrofuran was treated with excess methyl iodide. The mixture was stirred at 50 ° C. for 4 hours. The fine crystalline yellow solid (2.20 g, 94.1%) thus obtained had a melting point of 123-125 ° C. and was confirmed to be of the structure shown below.

Example 127 N- (2-fluoroethyl) -N '-[2- (1,4-dihydro-1-methyl-3-pyridinecarbonyloxy)
Ethyl] -N-nitrosourea Synthesis of quaternary salt type nitrosourea derivative 0.426 (1 mmol) prepared in Example 126 and 1-benzyl-1,2-dihydroisonicotinamide 0.21 g (1 mmol) in anhydrous ethanol 25ml
Was stirred at 0 ° C. under a nitrogen atmosphere for 4 hours. The solvent was evaporated under reduced pressure at about 30 ° C., the residue was suspended in chloroform, filtered and flash chromatographed on a short column of neutral alumina. Elution with chloroform provided the title compound in 55% yield. The product was assigned the following structure, which was consistent with the UV analysis.

Example 128 Synthesis of chloromethyl nicotinate In a suspension of 1.23 g (0.01 mol) of nicotinic acid in a mixture of 10 ml of water and 20 ml of tetrahydrofuran, 0.34 g (1 mmol) of tetrabutylammonium hydrogen sulfate and sodium bicarbonate were added.
19 g (0.038 mol) were added with vigorous stirring. Then, 1.81 g (0.011 mol) of chloromethyl chlorosulfate in 5 ml of tetrahydrofuran was added dropwise while maintaining the temperature at 30 ° C or lower. The reaction mixture was stirred for 1 hour, then separated, and the organic layer was dried by azeotropic distillation with acetonitrile / benzene 1: 1. The residue was passed through a column of neutral alumina and eluted with chloroform. The chloroform layer was evaporated to give 1.28 g (74.8%) of an oily residue.
This was confirmed by NMR analysis to be represented by the following structural formula.

Example 129 1- (pyridine-3-carbonyloxymethyl) -5
Synthesis of Fluorouracil 5-FU (1.31 g, 0.01 mol) was dissolved in 5 ml of dimethylacetamide and treated with 2.78 ml (0.02 mol) of triethylamine. 2.95 g (0.012 mol) of chloromethyl nicotinate in 5 ml of dimethylacetamide are added in one portion and the mixture is stirred for 24 hours, then filtered, washed with ethyl acetate,
Evaporated. The residue is chromatographed on a column of silica gel, first with benzene, then with benzene / chloroform 3: 1, then with benzene / chloroform 1: 1 and then with chloroform / benzene 3: 1, then with chloroform.
Finally, elution was carried out with chloroform / methanol 99: 1. Unreacted nicotinic acid ester elutes first, then the 1,3-bis isomer and finally the desired 1-isomer. This one
The isomer (1.3 g, 50%) has a melting point of 190-192 ° C.
It was confirmed by NMR analysis that it was represented by the following formula.

Example 130 1,3-Bis (pyridine-3-carbonyloxymethyl)
Synthesis of -5-fluorouracil 5-FU (1.31 g, 0.01 mol) was dissolved in 5 ml of dimethylacetamide and treated with 5.6 ml (0.04 mol) of triethylamine. After the addition of 6.8 g (0.04 mol) of chloromethyl nicotinate in 15 ml of dimethylacetamide in one portion, the mixture was stirred for 48 hours, filtered and the cake was washed with ethyl acetate. The filtrate was evaporated under reduced pressure, the residue was diluted with 100 ml of water and extracted with chloroform. The organic layer was evaporated and the residue was dried by azeotropic distillation with acetonitrile / benzene 1: 1. The residue was chromatographed on a column of neutral alumina, eluting in order with benzene, benzene / chloroform 1: 1 and benzene / chloroform / methanol 50: 50: 1 to give the bis-isomer 3.2 of the target compound.
g (80%) was obtained. It was confirmed by NMR analysis that this compound was represented by the following formula.

Example 131 3- (pyridine-3-carbonyloxymethyl) -5
Synthesis of Fluorouracil The 1,3-bis isomer prepared in Example 130 was dissolved in 10 ml of methanol and mixed with 20 ml of potassium carbonate / sodium bicarbonate buffer (0.1 M), pH 10.00. The mixture was stirred at room temperature for 2 hours, monitoring by thin layer chromatography showed that by then the bis isomer had completely disappeared. The resulting reaction mixture was evaporated and the residue was chromatographed on an alumina column, eluting sequentially with chloroform, chloroform / methanol 99: 1, and chloroform / methanol 96: 4. Collect each fraction,
When the fractions containing the desired 3-isomer are collected and evaporated, the title compound represented by the following structural formula has a melting point of 179-180.
C (0.2 g, 30%). The structure of this product was confirmed by NMR analysis.

Example 132 Synthesis of 1- (1-methyl-3-pyridiniumcarbonyloxymethyl) -5-fluorouracil iodide 1.93 g of the 1-isomer obtained in Example 129 was mixed with a sufficient amount of methyl iodide and acetonitrile. The resulting mixture was heated at reflux for 4 hours, then cooled and filtered to give 2.5 g of a pale yellow fluffy solid. The filtrate was evaporated and the residue was triturated with acetonitrile and filtered to give another 0.23 g of the expected product. Total yield was 2.73 g (92.25%). UV and NMR analysis confirmed that the product had the structure shown below.

Example 133 Synthesis of 1- (1,4-dihydro-1-methyl-3-pyridinylcarbonyloxymethyl) -5-fluorouracil 1 g of the product of Example 132 was cooled in an ice bath and degassed with argon. Dissolved in 20 ml of deionized water, then added 20 ml of ethyl acetate. The solution was stirred and to this was added 1.24 g of sodium bicarbonate, and after about 1 minute, 1.75 g of sodium dithionite. The reaction was allowed to proceed under an argon atmosphere and monitored by UV. After about 75 minutes, ethanol was added, the precipitated solid was filtered off, washed with water and methylene chloride, and dried under an argon atmosphere to give 400 mg of the title compound. UV and NMR analysis confirmed that the product had the structure shown below.

The separated aqueous layer was repeatedly extracted with chloroform, and combined with the ethyl acetate layer used for the reaction. The solid obtained after removal of the organic solvent was suspended in acetonitrile and filtered to give an additional 250 mg of product. 173-174
° C.

Example 134 Synthesis of 3- (1-methyl-3-pyridiniumcarbonyloxymethyl) -5-fluorouracil iodide Using the 3-isomer obtained in Example 131, the method described in Example 132 is repeated. The corresponding quaternary salt represented by the following formula can be obtained.

Example 135 Synthesis of 3- (1,4-dihydro-1-methyl-3-pyridinylcarbonyloxymethyl) -5-fluorouracil Using the product of Example 134, proceed as described in Example 133 By repeating, a corresponding dihydropyridine compound represented by the following formula can be obtained.

The pharmaceutical composition for parenteral administration used in the method of the present invention can be used for treating various conditions depending on the pharmacological properties of the drug selected for the purpose of administration. In the case of a redox carrier drug, the pharmacological properties of the drug substance itself used to derive the carrier drug will be the determinant. Regarding the carrier-type drug, for example, in one preferred embodiment, a redox system is derived from dopamine or L-dopa or a protected derivative thereof, and the obtained redox derivative / cyclodextrin mixture (inclusion compound) is used. The composition for parenteral administration contains
It can be used to elicit a sustained brain-specific dopamine-type (eg, anti-Parkinson's disease or anti-hyperprolactinemia) response in animals to which it has been administered. Similarly, any of the redox derivatives / diclodextrin-based parenteral drug compositions of the present invention derived from other centrally acting drugs can be obtained by supplying the drug substance itself to the brain. It can be used to elicit a type of pharmacological response. That is, when the centrally acting drug substance is an antitumor / anticancer drug, the parenterally administered drug composition of the present invention is used to elicit an antitumor / anticancer response; When a sympathomimetic is used, the pharmaceutical composition of the present invention is used to elicit a sympathetic stimulation or a response similar to amphetamine; The drug composition is used to elicit an anticonvulsant response; if the drug substance is a tranquilizer, the drug composition of the present invention is used to elicit a tranquilizer response; When a drug, the pharmaceutical compositions of the present invention are used such as to elicit an antidepressant response.

The pharmaceutical composition for parenteral administration used in the method of the present invention comprises β
-Or pharmacologically effective in aqueous solutions containing hydroxypropyl, hydroxyethyl, glucosyl, maltosyl or maltotriosyl derivatives of γ-cyclodextrin, preferably hydroxypropyl-β-cyclodextrin at a concentration of about 20-50%. It comprises an amount of at least one lipophilic and / or water-stable agent to be administered.

The composition is sterile and pyrogen-free, for example,
Edited by Alfonso R. Gennaro, Remington's Pharmaceutical S
ciences 17th edition (1985), pp. 1518-1552 (Mack Publishi
ng Co.). The aqueous sterile injectable solution may further contain additives such as antioxidants, buffers, bacteriostats and isotonicity regulators that are acceptable for parenteral administration. As is well known in the art, various single- and multi-use containers (eg, sealed ampules or biases) can be used.
The essential components of the composition for sterile parenteral administration, i.e., drug, water and certain cyclodextrin derivatives, can be prepared in a variety of ways, as long as the solution ultimately administered to the patient contains appropriate amounts of each of these essential components. Can be present in the composition. For example, the composition comprising the drug / cyclodextrin / water of the present invention can be provided as a single-use or multiple-use container so that it can be injected as it is. As another example, the concentrated drug / cyclodextrin / water solution can be provided in a separate container from the diluent liquid (water or cyclodextrin / water mixture). In this case, the contents of the two containers are mixed and used as a parenteral pharmaceutical composition at a concentration suitable for injection. As yet another example, a drug or drug / cyclodextrin mixture (or clathrate) may be lyophilized in one container and the dilution liquid (cyclodextrin in another container) in another container. Water or cyclodextrin / water mixture, depending on the amount). In this case as well, the contents of both containers are also mixed to prepare a composition containing the essential components at an appropriate concentration and then used. In each case, the contents of each container are sterile.

The therapeutic dose range for administering the drug in the composition for parenteral administration of the present invention will generally be the drug itself (or, if the drug is a carrier drug, the drug substance itself before binding to the carrier). Will be equal to or less than the dose normally used to administer (in some cases, much less). Of course, such a therapeutic dose range
It varies depending on conditions such as the species and weight of the patient, the administration conditions of the pharmaceutical composition, and the type of parenteral administration to be employed. The dosage of the parenterally administered pharmaceutical composition of the present invention required to provide a desired amount of the active ingredient will, of course, vary with the concentration of the drug in the composition.

While the invention has been described in terms of various preferred embodiments, it will be apparent to those skilled in the art that various modifications, substitutions, omissions, and changes may be made within the scope of the invention.

[Brief description of the drawings]

FIG. 1 shows that estradiol-CDS 17β-[(1
- methyl-1,4-dihydro-3-pyridinyl) carbonyloxy] estra-1,3,5 (10) - triene-3-ol (hereinafter E ₂ -CDS abbreviated) in dimethyl sulfoxide E ₂ - the CDS 15mg / kg (○) or after systemic administration to rats in an amount of E ₂ -CDS to 5 mg / kg of the state inclusion compounds of hydroxypropyl -β- cyclodextrin in water (△), lung tissue the concentration of E ₂ CDC in (left), and the same E
_2- After administration of CDS, the corresponding quaternary cation, 17β-
The lung concentration (right side) of [(1-methyl-3-pyridinium) carbonyloxy] estra-1,3,5 (10) -trien-3-ol (hereinafter abbreviated as E ₂ Q ⁺ or Quat) was determined as follows:
_{Μg [C B / D, μg} / g] per dose 1g were compared by correcting the amount administered as a pair of semi-log plot, FIG. 2, E ₂ -CDS rat in dimethylsulfoxide
To 15 mg / kg (□) or hydroxypropyl-β in water
- the state of inclusion compounds of cyclodextrin E ₂ -CDS
The 5mg / kg (■) after systemic administration of E ₂ Q ⁺ or concentration of Quat quaternary cation in the brain at a particular point in time, the dose
4 is a bar graph showing the dose corrected as ng per gram.

Claims (33)

(57) [Claims] 1. The method according to claim 1, wherein the lipophilic and / or water-labile drug is deposited in or near the injection site after parenteral administration and / or within the lung or other organ itself due to deposition within the lung or other organ itself. A method of reducing the tendency to collect in a parenteral administration aqueous solution containing about 20 to 50% of a hydroxypropyl, hydroxyethyl, glucosyl, maltosyl or maltotriosyl derivative of β- or γ-cyclodextrin. A method comprising: dissolving a composition. 2. The method according to claim 1, wherein the lipophilic and / or water-unstable drug is deposited at or near the injection site after parenteral administration and / or deposited within the lung or other organ itself. A method of reducing the tendency to collect in a parenteral administration aqueous solution containing about 20 to 50% of a hydroxypropyl, hydroxyethyl, glucosyl, maltosyl or maltotriosyl derivative of β- or γ-cyclodextrin. Wherein the aqueous solution is about 20 to 50
% Hydroxypropyl-β-cyclodextrin, the drug is a reduced form of a dihydropyridine pyridinium salt redox system for drug delivery to the brain, biooxidative, blood-brain barrier permeable, lipoidal A method which is other than a dihydropyridine compound. 3. The method according to claim 1, wherein said aqueous solution is a substantially isotonic solution. 4. The method according to claim 1, wherein the drug is an anti-neoplastic (anti-tumor) drug.
The method according to claim 1. 5. The method according to claim 1, wherein the drug is a sedative, a tranquilizer, an anticonvulsant, an antidepressant, a hypnotic, a muscle relaxant or a sedative. 6. The method according to claim 1, wherein the drug is an androgen, an estrogen,
The method according to claim 1 or 2, which is a progestin or an anti-inflammatory steroid. 7. The method according to claim 1, wherein the drug is a steroid hypnotic or an anesthetic. 8. The method according to claim 1, wherein the drug is an anticoagulant, a cardiotonic, a vasodilator, a vasoconstrictor, a platelet inhibitor or an antiarrhythmic. 9. The method according to claim 1, wherein the drug is an antifungal drug, an antigenic animal drug, an antibacterial drug, an antibiotic or an antiviral drug. 10. The method according to claim 1, wherein the drug is a vitamin / nutritional factor, an emetic, an antiemetic, a diuretic, a nonsteroidal anti-inflammatory drug, an anesthetic, a hypoglycemic drug, a radiological diagnostic drug, a carbonic anhydrase inhibitor,
Anesthetic antagonists, narcotics, narcotics-antagonists,
The method according to claim 1 or 2, which is a pharmacologically active protein, a dopaminergic / anti-Parkinson's disease drug, or a drug for treating Alzheimer's disease. 11. The method according to claim 1, wherein the drug is chlorambutyl, lomustine, melphalan, methotrexate, hexamethylmelamine, teniposide, etoposide, semustine, fazarabine, mercaptopurine, tubrazole, carmofur, carmustine, amsacrine, bruceantin, diaziquon, didemnin B, echinomycin. Or PCNU
5. The method of claim 4, wherein 12. The method according to claim 5, wherein the drug is a barbitur derivative or a benzodiazepine compound. 13. The method according to claim 5, wherein the drug is phenytoin, pentobarbital, phenobarbital, secobarbital, sulpiride, etomidate, chlordiazepoxide, diazepam, medazepam, oxazepam or lorazepam. 14. The method according to claim 11, wherein the drug is dexamethasone, hydrocortisone, prednisolone, 17β-estradiol,
The method according to claim 6, which is α-ethynyl estradiol, ethinyl estradiol 3-methyl ether, estriol, norethindrone, norethindrone acetate, norgestrel, ethisterone, medroxyprogesterone acetate, progesterone, 17-methyltestosterone or testosterone. 15. The method of claim 7, wherein said drug is alphaxalone. 16. The method according to claim 16, wherein the drug is dicoumarol, digoxin,
The method according to claim 8, which is digitoxin, nitroglycerin, flunarizine, alprostadil or prostacyclin. 17. The method of claim 9, wherein said agent is ampicillin, penicillin G, ketoconazole, itraconazole, metronidazole benzoate, miconazole, flubendazole or co-trimoxazole. 18. The method according to claim 18, wherein the drug is retinol, vitamin A acetate, cholecalciferol, retinal, vitamin E,
11. The method according to claim 10, which is D or K, apomorphine, chlorthalidone, furosemide, spironolactone, indomethacin, piroxicam, flurbiprofen, acetazolamide, lidocaine, acetohexamide, dimenhydrinate, L-dopa or THA. 19. The drug according to claim 1, wherein the drug is a biooxidative, blood-brain barrier permeable, lipoidal dihydropyridi compound which is a reduced form of a redox system of a dihydropyridinepyridinium salt type for drug delivery to the brain. Or the method of 2. 20. The method of claim 19, wherein said aqueous solution is a substantially isotonic solution. 21. The dihydropyridine compound represented by the general formula [D-DHC] wherein [D] is a centrally acting drug, and [DH]
C] is a biooxidative, blood-brain barrier-permeable, lipoidal reduced form of a dihydropyridinepyridinium salt-type redox carrier).
The described method. 22. The centrally acting drug may be a dopamine agonist, an androgen agonist, an anticonvulsant, an anxiolytic, a neurotransmitter, an antibiotic or an antibacterial, an antidepressant, an antiviral, or a drug. A cancer or an antitumor drug, an anti-inflammatory drug, an estrogen, or a progestin.
21. The method according to 21. (23) The centrally acting drug species is dopamine, testosterone, phenytoin, GABA, valproic acid, tyrosine, methicillin, oxacillin, benzylpenicillin, cloxacillin, dicloxacillin, desipramine, acyclovir (acycloguanosine), Fluorothymidine, zidovudine, hydroxy-CCNU, chlorambutyl, tryptamine, dexamethasone, hydrocortisone, ethinylestradiol, norethindrone, estradiol, ethisterone, norgestrel, estrone, estradiol 3-methyl ether, estradiol benzoate, norethinodrel, mestranol, indomethacin, NUproxen, NUPROXEN
23. The method according to claim 22, which is HENU or 5-FU. 24. A compound represented by the general formula [D-DHC], wherein 1-methyl-3- [N- {β- [3,4-bis (pivalyloxy) phenyl] ethyl} carbamoyl] -1,
4-dihydropyridine, 1-methyl-3- [N-Δβ-
[3,4-bis (isobutyryloxy) phenyl] ethyl}] carbamoyl-1,4-dihydropyridine, N-
{Β- [3,4-bis (pivalyloxy) phenyl] ethyl} aminocarbonyloxymethyl-1,4-dihydro-
1-methyl-3-pyridinecarboxylate, 17β-
[(1,4-dihydro-1-methyl-3-pyridinylcarbonyl) oxy] androst-4-en-3-one,
17β-{[(3 ″ -carbamoyl-1 ′, 4′-dihydropyridinyl) acetyl] oxy} androst-4-en-3-one, 5,5-diphenyl-3-[(1′-methyl -1 ', 4'-dihydropyridin-3'-yl) carbonyloxymethyl] -2,4-imidazolidinedione,
-[(3'-carbamoyl-1 ', 4'-dihydropyridin-1'-yl) acetyloxymethyl] -5,5-diphenyl-2,4-imidazolidinedione, 3- [3'-
(3 "-carbamoyl-1", 4 "-dihydropyridine-
1 "-yl) propionyloxymethyl] -5,5-diphenyl-2,4-imidazolidinedione, 1-methyl-3
-N- [3- (benzyloxycarbonyl) propyl]
Carbamoyl-1,4-dihydropyridine, 1-methyl-
3- {N-[(3'-cyclohexylcarbonyl) propyl]} carbamoyl-1,4-dihydropyridine, 1-
Methyl-3- [2 '-(2 "-propyl) pentanoyloxy] ethylcarbamoyl-1,4-dihydropyridine, 1-methyl-3- [2'-(2" -propyl) pentanoyloxy] ethoxycarbonyl- 1,4-dihydropyridine, 1- [2 '-(2 "-propyl) pentanoyloxy] ethyl-3-carboxamide-1,4-dihydropyridine, 1-methyl-3- {N-[(1'-ethoxycarbonyl) ) -2 '-(4 "-Pivaloyloxyphenyl) ethyl] {carbamoyl-1,4-dihydropyridine, 1-methyl-3- {N-[(1'-ethoxycarbonyl) -2'-(4" -Isobutyryloxyphenyl)
Ethyl]｝ carbamoyl-1,4-dihydropyridine,
[[(1,4-dihydro-1-methyl-3-pyridinyl)
[Carbonyl] oxy] methyl • [2S- (2α, 5α, 6
β)]-3,3-dimethyl-7-oxo-6-[(2,6-dimethoxy) benzamido] -4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylate,
[[(1,4-dihydro-1-methyl-3-pyridinyl)
[Carbonyl] oxy] methyl • [2S- (2α, 5α, 6
β)]-3,3-dimethyl-6- (5-methyl-3-phenyl-4-isoxazolecarboxamide) -7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylate , [[(1,4-dihydro-1-
Methyl-3-pyridinyl) carbonyl] oxy] methyl. [2S- (2α, 5α, 6β)]-3,3-dimethyl-7-oxo-6-[(phenylacetyl) amino] -4-thia-1- Azabicyclo [3.2.0] heptane-2-carboxylate, [[(1,4-dihydro-1-methyl-3-pyridinyl) carbonyl] oxy] methyl. [2S- (2
α, 5α, 6β)]-6- [3- (2-chlorophenyl)-
5-methyl-4-isoxazolecarboxamide]-
3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylate,
[[(1,4-dihydro-1-methyl-3-pyridinyl)
[Carbonyl] oxy] methyl • [2S- (2α, 5α, 6
β)]-6- [3- (2,6-dichlorophenyl) -5
Methyl-4-isoxazolecarboxamide] -3,3
-Dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylate, [｛N-
[3- (10,11-dihydro-5H-dibenzo [b, f] azepin-5-yl)] propyl-N-methylamino {carbonyloxy] methyl-1,4-dihydro-1-methyl-3
-Pyridinecarboxylate, [1- {N- [3- (1
0,11-dihydro-5H-dibenzo [b, f] azepine-5-
Yl)] propyl-N-methylamino {carbonyloxy] ethyl-1,4-dihydro-1-methyl-3-pyridinecarboxylate, 1-methyl-3-{[2- (9-
Guanylmethoxy) ethoxy] carbonyl {-1,4-dihydropyridine, 3 '-(1,4-dihydro-1-methyl-3-pyridinylcarbonyl) -5'-pivaloyltrifluorothymidine, 3'-azido- 3'-deoxy-
5 '-[(1-methyl-1,4-dihydro-3-pyridinyl) carbonyl] thymidine, N- (2-chloroethyl)
-N '-[4- (1,4-dihydro-1-methyl-3-pyridinecarbonyloxy) cyclohexyl] -N-nitrosourea, N- (2-fluoroethyl) -N'-[2-
(1,4-dihydro-1-methyl-3-pyridinecarbonyloxy) ethyl] -N-nitrosourea, N- (2-chloroethyl) -N '-[2- (1,4-dihydro-1-methyl- 3-pyridinecarbonyloxy) ethyl] -N-
Nitrosoureas, 1-methyl-3-[(N- {2- [4-
({4- [bis (2-chloroethyl)] amino {phenyl) butanoyloxy] ethyl}) carbamoyl] -1,
4-dihydropyridine, 1-methyl-3- (N- {4-
[4- (4-{[bis (2-chloroethyl)] amino}
Phenyl) butanoyloxy] cyclohexyl {carbamoyl) -1,4-dihydropyridine, 1-methyl-3-
[(N- {2- [4-({4- [bis (2-chloroethyl)] amino} phenyl) butanoyloxy] propyl}) carbamoyl] -1,4-dihydropyridine, 1-
Methyl-3 [(N- {2-phenyl-2- [4-({4
-[Bis (2-chloroethyl)] amino {phenyl) butanoyloxy] {ethyl) carbamoyl] -1,4-dihydropyridine, 1-methyl-3- [N-({1- [4
-(4-{[bis (2-chloroethyl)] amino} phenyl) butanoyloxy] cyclohexyl} methyl) carbamoyl] -1,4-dihydropyridine, 1-methyl-
3-N- [2- (3-indolyl) ethyl] carbamoyl-1,4-dihydropyridine, 9-fluoro-11β, 17-
Dihydroxy-16α-methyl-21-｛[(1-methyl-
1,4-dihydropyridin-3-yl) carbonyl] oxy} pregna-1,4-diene-3,20-dione, 11β, 17-
Dihydroxy-21-{[(1-methyl-1,4-dihydropyridin-3-yl) carbonyl] oxy} pregn-
4-ene-3,20-dione, 3-hydroxy-17β-
[(1-methyl-1,4-dihydropyridin-3-yl)
Carbonyl] oxyestra-1,3,5 (10) -triene, 3-hydroxy-17β-{[(1-methyl-1,4-
Dihydropyridin-3-yl) carbonyl] oxy}-
19-nor-17α-pregna-1,3,5 (10) -triene-2
0-in, 3-[(1-methyl-1,4-dihydro-3-pyridinyl) carbonyloxy] estra-1,3,5 (10)
-Trien-17-one, 17β-[(1-methyl-1,4-
Dihydro-3-pyridinyl) carbonyloxy] estra-1,3,5 (10) -trien-3-ol-3-methyl ether, 3,17β-bis-{[(1-methyl-1,4-dihydropyridine- 3-yl) carbonyl] oxy {estra-1,3,5 (10) -triene, 3- (phenylcarbonyloxy) -17-β-} [(1-methyl-1,4-dihydropyridin-3-yl) Carbonyl] oxydiestra-1,3,5 (10) -triene, 3-methoxy-17β-
{[(1-methyl-1,4-dihydropyridin-3-yl) carbonyl] oxy} -19-nor-17α-pregna-1,3,5 (10) -trien-20-yne, 17β-{[( 1
-Methyl-1,4-dihydropyridin-3-yl) carbonyl] oxy} -19-norpregn-4-en-20-yn-3-one, 17β-{[(1-methyl-1,4-dihydropyridin-3 -Yl) carbonyl] oxy} pregn-4-en-20-yn-3-one, 13-ethyl-17β-
{[(1-methyl-1,4-dihydropyridin-3-yl) carbonyl] oxy} -18,19-dinolpregne-
4-en-20-yn-3-one, 17β-{[(1-methyl-1,4-dihydropyridin-3-yl) carbonyl]
Oxy} -19-norpregn-5 (10) -en-20-yn-3-one, 1-methyl-3- [N- (2- {1-
(P-chlorobenzoyl) -5-methoxy-2-methyl-3-indolyl] acetoxy {ethyl) carbamoyl] -1,4-dihydropyridine, 1-methyl-3- {N
-[2- (6-Methoxy-α-methyl-2-naphthalenylacetoxy) ethyl] carbamoyl} -1,4-dihydropyridine, 3- (1,4-dihydro-1-methyl-3-
Pyridinylcarbonyloxymethyl) -5-fluorouracil, or 1- (1,4-dihydro-1-methyl-3)
24. The method according to claim 23, which is -pyridinecarbonyloxymethyl) -5-fluorouracil. 25. The aqueous solution of claim 1, wherein the aqueous solution contains about 20-50% hydroxypropyl-β-cyclodextrin.
19. The method according to any one of claims 18 to 18. 26. The drug is a biooxidative, blood-brain barrier permeable, lipoidal dihydropyridine compound which is a reduced form of a dihydropyridinepyridinium salt type redox system for drug delivery to the brain, and wherein the aqueous solution is 20 ~
The method according to claim 1, wherein the method contains 50% hydroxypropyl-β-cyclodextrin. 27. An improved non-human animal having reduced propensity to collect in the lung or other organs after parenteral administration of the drug due to deposition at or near the injection site and / or within the lungs or other organs themselves. A pharmaceutical composition for oral administration, wherein the composition comprises a hydroxypropyl, hydroxyethyl, glucosyl, maltosyl or maltotriosyl derivative of β- or γ-cyclodextrin.
A drug composition for parenteral administration, characterized in that a drug that is lipophilic and / or unstable in water is dissolved in an aqueous solution containing 20 to 50%. 28. An improved non-human animal having reduced propensity to collect in the lungs or other organs following parenteral administration of the drug due to deposition at or near the injection site and / or deposition within the lungs or other organs themselves. A pharmaceutical composition for oral administration, wherein said composition is lipophilic and / or unstable in water containing about 20-50% hydroxypropyl, hydroxyethyl, glucosyl, maltosyl or maltotriosylcyclodextrin. Characterized in that, when the aqueous solution contains about 20 to 50% hydroxypropyl-β-cyclodextrin, the drug is supplied to the brain by drug delivery. Biohydrogenation, blood-brain barrier permeability, which is a reduced form of dihydropyridine pyridinium salt redox system for
A pharmaceutical composition for parenteral administration, which is other than a lipoidal dihydropyridine compound. 29. The pharmaceutical composition for parenteral administration according to claim 27, wherein the aqueous solution is a substantially isotonic solution. 30. The pharmaceutical composition for parenteral administration according to claim 27 or 28, wherein the drug is one according to any one of claims 4 to 18. 31. The pharmaceutical composition for parenteral administration according to claim 27 or 28, wherein said drug is one according to any one of claims 19 and 21 to 24. 32. The pharmaceutical composition for parenteral administration according to any one of claims 27 to 30, wherein said aqueous solution contains about 20 to 50% hydroxypropyl-β-cyclodextrin. 33. The drug is a biooxidative, blood-brain barrier-permeable, lipoidal dihydropyridine compound which is a reduced form of a dihydropyridinepyridinium salt-type redox system for drug delivery to the brain, and wherein the aqueous solution is 20 ~
28. The pharmaceutical composition for parenteral administration according to claim 27, which contains 50% hydroxypropyl-β-cyclodextrin.