TWI282783B - Disodium salts, monohydrates, and ethanol solvates for delivering active agents - Google Patents

Abstract

The inventors have discovered that the disodium salt of certain delivery agents has surprisingly greater efficacy for delivering active agents than the corresponding monosodium salt. Furthermore, the inventors have discovered that the disodium salts of these delivery agents form solvates with ethanol and hydrates with water. The delivery agents have the formula, wherein R1, R2, R3, and R4 are independently hydrogen, halogen, C1-C4 alkyl, or C1-C4 alkoxy; and R5 is a substituted or unsubstituted C2-C16 alkylene, substituted or unsubstituted C2-C16 alkenylene, substituted or unsubstituted C1-C12 alky (arylene), or substituted or unsubstituted aryl (C1-C12 alkylene). The hydrates and solvates of present invention also have surprisingly greater efficacy for delivering active agents, such as heparin and calcitonin, than their corresponding monosodium salts and free acids. The present invention provides an alcohol solvate, such as ethanol solvate, of a disodium salt of a delivery agent of the formula above. The invention also provides a hydrate of a disodium salt of a delivery agent of the formula above. Preferred delivery agents include, but are not limited to, N-(5-chlorosalicyloyl)-8-aminocaprylic acid (5-CNAC), N-(10[2-hydroxybenzoyl]amino)decanoic acid (SNAD), and sodium N-(8-[2-hydroxybenzoyl]amino)caprylate (SNAC). The invention also provides methods of preparing the disodium salt, ethanol solvate, and hydrate and compositions containing the disodium salt, ethanol solvate and/or hydrate.

Description

1282783 V. INSTRUCTIONS (1) The present invention claims US Patent Application Serial No. 60/127754 (filed on Apr. 5, 999), and U.S. Patent Application Serial No. 60/1861 43 (March 1, 2000) Application, US Patent Application Serial No. 60/1861 42 (filed on March 1, 2000) and U.S. Patent Application Serial No. 60/1 9 1 2 86 (filed March 21, 2000). FIELD OF THE INVENTION This invention relates to a disodium salt of a delivery agent, such as N-(5-chiorosaiicyioyi) \ - 8-aminocapryiic acid N-(10-[2-hydroxybenzomethyl]amino)decanoic 丨acid) or N-(8-[hydroxyloxy) N-(8-[2-hydroxybenzoyi]amino)capryiic acid), a species j

An alcohol solvate of the disodium salt and an I monohydrate as a disodium salt for transferring an active agent and a process for the preparation thereof. BACKGROUND OF THE INVENTION The composition of active agents for oral delivery, such as heparin and calcitonin, with modified amino acids, is disclosed in U.S. Patent Nos. 5,773,647 and 5,866,536. For example, 丨~(5~ gasified salicylate)_8-aminooctanoic acid (5-CNAC), N_(10-[2-hydroxyoxyindolyl]amino)decanoic acid (SNAD) or N- (8 - [2-Hydroxyphenylphenyl]amino)octanoic acid (SNAC). Many current formulations contain an active agent, such as hepatic stagnation and about modulating hormone, which are transmitted via routes other than the oral cavity. Oral delivery is better than other methods, and it can increase the acceptance of the disease. ,,~stupid

1282783 V. INSTRUCTIONS (2) ! Therefore, it is necessary to improve the oral administration of active agents such as hepatic cadherin I and calcium regulating hormones. BRIEF SUMMARY OF THE INVENTION The inventors have revealed that the disodium salt of certain specific delivery agents has a surprisingly good effect on the delivery of the active agent over the corresponding monosodium salt. In addition, the inventor

I found that the disodium salt of such a delivery agent forms a solvate with an alcohol or forms an I hydrate with water. This transfer agent has the following sound pattern: ! I 1 R4 Ο

Wherein: R1, R2, R3, and R4 are each independently hydrogen, -OH, -NR6R7, halo, alkyl or [alkoxy (alkoxy)]. R5 is substituted or unsubstituted C>-C6 alkylene, substituted or unsubstituted |

(S2-(:6 alkenylene), substituted or unsubstituted [c2-csalkyl(arylene)]f or substituted or unsubstituted [aryl (C2 - C6 sub-hospital) )]; and ! ! R6 and R7 may each be hydrogen 'oxygen or α-(:4 alkyl. | I The hydrate or solvate of the present invention is equivalent to the corresponding monosodium salt or the free acid, for delivery Active agents such as hepatic cadherin and calcium regulate 'hormone, also i

1071-3452-PF.ptd Page 10 1282783 V. Description of invention (3) It has amazingly good results. * ϋ | - The present invention provides an alcohol of the disodium salt of the above-mentioned structural formula| / phenolic compound such as methanol, ethanol, propanol, propylene glycol (pr〇py i ene jg 1 yco i) and other hydrogen Solvate of an oxy group. According to one of the preferred embodiments, the pseudoalcohol/solvate is an ethanol solvate. The present invention also provides a monohydrate of a monohydrate such as the above-mentioned chemical formula. Preferred transfer agents include, but are not limited to, N-(5-chlorosulphate)-8-amino group in acid (5-CNAC), Ν-(1〇-[2-hydroxybenzophenone Amino) decanoic acid j (SNAD), N-(8 - [2-hydroxyphenylphenyl]amino)octanoic acid (SNAC), | 8-(N - 2-hydroxyoxy-4- 8-(N-2-hydroxy-4-methoxybenzoyi)aminocarpryiic acid, such as the compound 67 in the U.S. Patent No. 5,773,647), a monohydroxybenzoinyl) amine group N-(9-2-hydroxybenzoyi)aminononaic acid or 9-salicyl〇ylaminononaic. The present invention also provides a process for producing the disodium salt of the present invention by drying the ethanol solvate of the present invention. According to a preferred embodiment, the ethanol solvate is prepared by the following method. Another embodiment of the invention is a process for making the ethanol solvate of the invention. The method comprises (a) dissolving a transfer agent of the above formula in ethanol to form a transfer agent/ethanol solution; (b) reacting the transfer agent/ethanol solution with a molar excess of the sodium-containing salt to form an ethanol solvate. Another embodiment of the invention is a method of making a hydrate of the invention. j The method comprises (a) obtaining an ethanol solvate (13) of the disodium salt of the transfer agent.

1071-3452-PF.ptd Page 11 1282783 I. V. INSTRUCTIONS (4) ?

The solvate is dried to form an anhydrous di-nano salt, and (C) the anhydrous di-nano salt is hydrated to form a hydrate. Another embodiment of the invention is a composition of a disodium salt comprising a transfer agent

5 I j things. i Another embodiment of the invention is a composition comprising at least one of the two sodium salts, ethanol solvates or hydrates of the invention, and at least one active 丨-I ^ | agent. Preferred active agents include, but are not limited to, 5 heparin and calcium modulating stimuli

Prime. The composition may be formulated in a pharmaceutically acceptable form, for example, as an oral pharmaceutically acceptable form. Another embodiment of the present invention is a method of administering an active agent to a desired emollient, which comprises The animal is administered a composition of the invention. (- I) As used herein, the term "substituted" includes, but is not limited to, the following substituents: halogen, hydroxide, C4 alkyl or Ci-C4 alkoxy (alkoxy)i Any one or any of its components. i

The "alkyl alkoxy" alkylene group, "alkenylene" "II alkyl (arylene)" and "alkyl (alkylene)", respectively, are used, but are not limited to, 丨i linear Or a branched alkyl group, an alkoxy group, a pyridinylene group, an s-dense group, a pyridyl group (arylene group) π and an 'alkyl group (alkylene group). i Disodium Salt | The disodium salt can be prepared from a solvate of ethanol by volatilizing or drying the ethylene glycol therein by conventional techniques to form an anhydrous di-nano salt. In general, |j drying is carried out at a temperature of 80 to 120 ° C, preferably at 85 to 90 ° C (the optimum temperature is about 85 ° C. A representative drying step is 26 j ying mercury column or higher pressure. Anhydrous disodium salt usually contains less than j -

1071-3452-PF.ptd Page 12 1282783 5% by weight of ethanol, while the latter contains less than 2% by weight of ethanol (based on 100% anhydrous di-n-salt total weight) 1 Transmitter 2 The sodium salt can also be made into a slurry of 3 kinds of sodium alkoxide by adding a transfer agent in water to a slurry and adding a 2 molar equivalent sodium hydroxide solution, sodium alkoxide or the like. , / qndiu ffi methoxide), B": including, but not limited to, sodium decoxide <s0Q 1 ^ j sodium (s 〇diumeth ο X ide ) and its 'heart. Another method of 't t to make two sodium salts is to react the transfer agent with a molar equivalent of sodium hydroxide to form a single sodium salt for the transfer, and then add an additional

And, a substance is transferred to a C I-mol equivalent of sodium hydroxide to form a mono-sodium salt. The solution containing the disodium salt can be concentrated by vacuum distillation to form a sticky paste to separate the solid. The paste was dried in a vacuum: box to give the disodium salt of the transfer agent of the i solid. It is also possible to dry the disodium-producing water-cooled spray to obtain solids, for example, in the U.S. Patent Law. The method of the present invention is based on the method of the prior art, and the other aspect of the present invention proposes a group of more than 20%. The delivery agent disodium salt, preferably a minimum of 60% of the delivery agent disodium salt (based on 100% of the total weight of the composition of the delivery agent and its salts). According to a specific embodiment, the composition comprises at least about 10, 30, 40, 50, 70 or 80% by weight of the carrier disodium salt (based on 1% by weight of the total weight of the composition) Transfer agent and its salts). More preferably, the composition contains at least about 90% of the transaminant disodium salt (based on the total weight of the composition in the composition of 1 〇 〇 % of the transfer agent and its salts). ’

1071-3452-PF.ptd Page 13 1282783

V. Description of invention (6)

The best is that the composition contains a large amount of the pure sodium salt of the tanning agent. Here, the term "a large amount of pure." is used to mean that the non-di-nano salt material in the transfer agent composition is less than 4% by weight, preferably less than 2% (base | in the composition i 〇n% of the total weight of the transfer agent and its salts). I

The term "ethanol solvate" as used herein includes, but is not limited to, I molecules or ionic complexes comprising molecules or ions of the molecule or ion and the carrier disodium strontium. A typical ethanol solvate comprises an ethanol molecule or ion in each molecule of the carrier-sodium salt. i 偻 = 7 sodium? The ethanol solvate can be prepared in the following manner: Zhai ♦ 50 ΐ 2 5 ethanol. Generally, every 1 gram of the transfer agent is dissolved in about 1 I 丨: in the pen-opened ethanol, the action ^ ^ f ^ / good 疋 dissolved in 2 to 10 ml of gt Φ. Ran | after the transfer agent / ethanol solution fish One by one, the salt of monosodium is reacted with the sodium salt of the sputum and the sputum, for example, %丨j is added to the ratio of the bismuth, which is more than the mutual enthalpy, such as: per-mole transfer agent Then, a single sodium salt-containing spoon = sub-s. This reaction produces an ethanol solvate. Suitable for, for example, formazan:: one is limited to 5 虱 emulsified sodium, burnt gas, preferably. Minimum = sodium oxyhydroxide and any mixed sterol solution of the foregoing, such as: 々 two molar equivalents of monosodium containing salts added | Sub. In general, this counter-reagent corresponds to a minimum of two moles of sodium, such as the reflux temperature of the dragon π π conjugate or the lower volatility" to &gt; The compound can be butyl.

1071-3452-PF.ptd Page 14 Add, sodium oxide to the transfer agent is recovered by conventional methods. For example, because the concentrate is concentrated. The slurry produced by the concentrated enthalpy can be steamed through the atmosphere _ ___ and can not be cooled again, and the solid product is recovered through the filtration of 1282783 j. The filtered cake, such as the filtrate, can be dried under vacuum to give the ethanol solvate. Hydrate!; | The term "hydrate" as used herein includes, but is not limited to, (i) a substance that contains water in the form of a molecule, and (ii) a crystalline form; One or more crystalline water molecules or a j-crystalline material containing free water. The composition containing the disodium salt hydrate preferably contains at least about I 80% by weight, more preferably at least about 90%, most The best is about 95% of the bismuth hydrate of the disodium salt (based on 100% of the total weight of the disodium salt hydrate in the composition). Hydrate can be dried by drying the ethanol solvate. The second salt, i is prepared by combining anhydrous di-n-salt brine. Preferably, it is a monohydrate forming a di-salt salt. Since the anhydrous disodium salt is very hygroscopic, the hydrate is exposed to the humidity of the large j gas. In general, the step of hydration is carried out at room temperature to 5 (left to right between TC, and in an environment with a relative humidity of at least 50%. Preferably, the step of hydration is between room temperature and 30 ° C. For example, the step of hydration can be carried out at 40 ° C and a relative humidity of 75%. The anhydrous disodium salt may also optionally be hydrated with steam. According to a preferred embodiment, the drying and hydrating steps may be carried out in an oven. Preferably, the material is not exposed to the atmosphere until both steps are completed. , a composition of an ethanol solvate and a hydrate, and a pharmaceutical dosage form thereof. The invention also provides a composition, such as a pharmaceutical composition, comprising at least one of the disodium salt, ethanol solvate or hydrate of the invention

1071-3452-PF.ptd Page 15 1282783 V. INSTRUCTIONS (8) Sex agents. A representative composition of the present invention is a disodium salt, an ethanol solvate and/or a hydrate of the present invention comprising an effective amount of one or more of the present invention, such as an effective amount of a disodium salt, an ethanol solvate and/or Or a hydrate to deliver the desired effect by delivering an active agent. I The active agents suitable for use in the present invention include biological active agents and chemical agents, active agents, package #, but are not limited to, insecticides, pharmaceutical preparations, and therapeutic agents. i For example: a biological or chemical active agent suitable for use in the present invention includes

But not limited to, proteins, peptides, peptides, hormones, polysaccharides, and I, especially sticky sf lava, carboniferous water compounds, lipids, other organic matter, and especially for themselves cannot pass (or In the dose, only one part of the drug can be used to win the gastric mucosa, and/or it is easy to be acid or enzyme in the gastrointestinal tract |

丨 chemically cracked component ' or any mixture thereof. I

Further examples of II include, but are not limited to, synthetic, natural, or a mixture of two kinds of growth hormones, including human growth hormones (hGH), recombinant human growth hormones (rhGH) , cattle growth hormone, | and growth hormone hormone release hormone hormone (releasing hormone), interferon, including alpha, 10,000 and r interferon, interleukin l ( Interleukin-1), white blood cells? Tengdaosu, containing mixed insulin, bovine, human and human, can choose the ion (counter i〇n) including sodium, zinc, calcium and ammonium ions, pancreatic ~ heart growth factor, including IGF-1, liver system Lectin, including isolated liver, j, = heparinoids, dermatans, low 卞 $ liver condensate, very low molecular weight hepatic condensate plate over August

1282783 V. INSTRUCTIONS (9) Hepatic calcin, I bow regulating hormone, including baby fish, 鳗 'fish, pig and human, erythoprotein, atrial nati!retie factor ' antigen, monoclonal antibody ( Monoclonal antibody) 'somatostatin, protease inhibitor, adrenocorticotropin, gonadotropin releasing hormone, oxytocin, luteal promoting hormone release (lentinizing — hormone — reieasing-hormone), stimulating, and follicle stimulating hormone, glucocerebrosidase, thrombopoietin, filgrastim, ProstagUndins 'cyclosporin, vasopressin, cromolyn sodium (sodium or 1 disodium chromoglycate), pan gram Vancomycin, demi err ioxamine (DFO), 畐4 甲甲甲〔 [parathyroid hormone (ΡΤΗ), including its fragments, Antibacterial agents, including antifungal agents, vitamins, analogs of such ingredients, fragments, mimetics or their polyethylene glycol (PEG) iodified derivatives, or mixtures thereof. Preferred active agents include, but are not limited to, hepatic statins and calcium regulating hormones. The amount of active agent in the composition is an effective amount to achieve the intended purpose. The amount in the composition is a pharmaceutically, biological, therapeutic or chemical effective amount. However, when a plurality of compositions are administered, the dose can be lowered. For example: ! The total effective dose is the cumulative value of all doses. When the composition is, the active agent is slow

1071-3452-PF.ptd Page 17 1282783 I V. INSTRUCTIONS (10) } | In slow release, the effective amount of active agent can also be higher than a therapeutic, biological, therapeutic or antispasmodic effective amount. . Such a composition generally has a sustained release coating which allows for a prolonged release of a pharmaceutical, biological, therapeutic or chemical active agent in the composition. |

The total amount of active agent applied to the sputum can be determined by those skilled in the art. However, the composition can deliver the activity more efficiently than the conventional composition, so that the target can be administered with a dose of less than the conventional dose or delivery system. While still maintaining the same blood concentration and / or therapeutic effect. i 1 According to a preferred embodiment, the composition comprises a carrier salt of a sodium salt and a calcium regulating hormone. Preferably, the delivery agent is 5-CNAC. Generally, the weight ratio of the calcium-regulating hormone to the disodium salt of 5-CNAC is determined according to the animal to be administered to the composition. For example, when the above composition is to be administered to a human, the weight ratio is from about 1:30 G to about 1:700, preferably 1: I' ί | 500 ^ for primates, weight The ratio is generally between 1:100 and 1: : j 500 or so. [The composition of the present invention may be in a liquid or solid form. Preferably, | i contains the composition of the disodium salt or/and hydrate of the present invention in a solid form. The composition may further comprise an additive, but is not limited to, a pH adjuster, an anti-septic, a flavoring agent, a flavoring agent, a fragrance, a wetting agent, a strongening agent, a coloring agent, a surfactant, a plasticizer, and a lubricating agent. A pharmaceutical carrier, a co-solvent, a ii excipient, a diluent, a disintegrating agent, or a combination of any of the above components. Dosing carriers suitable for 丨 1 ! I include, but are not limited to, water 'buffered phosphate, 1,2-propane||alcohol, ethanol, eucalyptus oil, 25 % propylene glycol aqueous solution, and the above ingredients |

i arbitrarily composed. Other additives include buffered phosphate 'lemon' acid, ethylene II

1071-3452-PF.ptd Page 18 1282783 V. INSTRUCTIONS (11) Good Alcohols and other dispersants. Stable additives can be added to the solution at concentrations ranging from 0.1 to 20% (w/v). The composition may further comprise one or more enzyme inhibitors such as j actinonin or epiactinonin and derivatives thereof. Other enzyme inhibition assays include, but are not limited to, a pulse jet (a p r 〇 t i n i η, r a s y i 〇 1) and a B o w m a π - B i r k inhibitor. i ' l .

The composition of the present invention can be prepared by mixing a disodium salt, a hydrate, and/or an ethanol-soluble compound, an active agent, and other selected additives in a dry mixture or in a solution of j. The mixture can be heated and/or converted slowly!

In order to enhance the dispersion effect of the above components in the solution. I The composition of the present invention may be formulated into a form of administration, particularly orally, in a pharmaceutical form, including, but not limited to, a capsule, a lozenge or a microparticle, such as a powder or microcapsule. i' According to a preferred embodiment, the dosage form can be a solid dosage form comprising a minimum of one of the disodium salt of the invention, an ethanol solvate, or a hydrated frozen j dry mixture with a minimum of one active agent. j

The above &quot;freeze-dried mixture: includes, but is not limited to, a mixture which is obtained by rapid cooling and dehydration. A representative hydrophobic solution is carried out in a vacuum in the case of freezing. The lyophilized mixture generally has no water damage on the large body, preferably based on the total weight of the composition 丨〇 , and the water content φ is less than 4%. I,, such a solid, administered dosage form can be prepared in the following manner: (A solution of a solution containing a plant above a transfer agent and one or more active agents; (b) j freeze the above solution to obtain freeze-drying Mixture; and (c) the cold j 1

1282783 V. INSTRUCTIONS (14) The device is cooled and the upper valve reactor is set to distillation. 4 1 · 6 | j kg of dry anhydrous sodium carbonate was injected into the reactor, and then the injection | |孑L was turned off. At this point, be careful of exhaust and a small amount of exothermic reaction. Will 7 7. 5 liters of dry |

I dry 6-chloro-2H-1,3-stupin-2, 4(3ίί)_dioxene was added to the reactor. And I

i I! Quickly dry 88 kg of ethyl-8-bromonate! ί \ ethyl (ethyl-8-bromooctanoate) was injected into the reactor. Reaction emptying to 22 |

i c I

|Up to 24 inches of mercury, the reactor temperature is increased to 65 to 75C. The reactor temperature j | is maintained in this temperature range and notes whether the contents are foaming. Reactor mixing j I was sampled and monitored by a gas chromatograph analyzer for its brominated ester (bromo I

^ I eater) A transition that disappears in the reaction mixture. The reaction was completed after 7 hours of ▲ I (0.6% of the brominated ester remained). After breaking the vacuum, the contents of the reactor were cooled to 45 to 501. The contents are passed through, and the filter is placed in a 2nd glass-lined 200-gallon reactor. U9 liters of ethanol (containing | | 〇 · 5 % of the 200-degree denatured alcohol) was injected into the first 200-gallon reactor | j, and the temperature was raised to 45 t. The pellet after centrifugation was washed with warm ethanol and the cleaning solution was poured into the reaction mixture of the second 200 gallon reactor. j The agitator in the second 200-gallon reactor is started. Reactor contents! The temperature is lowered to approximately 29 °C. 1 2 0 liters of deionized water was slowly added to the 2nd reactor | and water was poured directly into the contents. The reactor contents are then cooled to about 丨:. The intermediate was precipitated from the solution and maintained for about 9.5 hours. The resulting ‘ | mud aggregate is centrifuged. A 70 liter A release was injected into the reactor, the temperature j j was lowered to about 8 ° C, and the pellet after centrifugation was washed. The moist mass j | is placed in a double-layered polyethylene bag and placed in a paper-lined drum. Production | | Out of 123.5 kg of ethyl 8-(6-chloro-2H-1,3-alum 哄u(3H)_ two j

1071-3452-PF.ptd Page 22 1282783

I 嗣) octoate (ethyl 8-(6-cM〇ro-2H-;[,3-b0nz〇Xazine _2 4(3H)~dionyl)octanoate). 400 liters of pure water, USP and 45.5 kg of sodium hydroxide pellets were injected into a glass-lined 200-gallon reactor, and the stirrer was set at 1 〇〇 to tZOipni&gt;τγ of ethyl 8~ (g - gas - clumsy - 9 4 (3H) - diterpene) octanoate moist cake was also injected into the reactor and the injection port was closed. The cold clock water is injected into the condenser, and the valve reactor above it is set to atmospheric distillation. The reactor contents were heated to approximately 98 〇c and their transitions were monitored by HPLC. The reactor was at 68 for about 40 minutes from the start. The underflow was refluxed. However, after the ethanol was removed by distillation (after about 3 hours), the reaction temperature was raised to about 98 art. After the HPLC measurement, the starting material disappeared after about 4 hours. The reactor contents were cooled to about 27 t. 15 liters of distilled water was placed in a 2 〇 0 gallon reactor with a glass lining adjacent to USP 1981; the stirrer was set at 100 to 125 rpm. 1 〇 4 liters of concentrated hydrochloric acid (12M) was injected into the reactor and cooled to about 24 °C. The saponification reaction mixture was slowly injected (about 5 hours) into the glass-lined 200 gallon reactor. Due to the production of carbon dioxide, the material is divided into 45 liters and 45 liters, respectively, into two 2 gal gallons of reactor. The product then precipitates out of solution. The reaction mixture was adjusted to a pH of 2.0 to 4.0 with a 50% sodium hydroxide solution (2 kg of sodium hydroxide dissolved in 2 liters of water). The contents of the reactor were cooled to between 9 and 15. The intermediate gradually precipitated from the solution over 9 hours. The slurry of the reaction is centrifuged to separate the intermediate. 5 liters of pure water was injected into the glass liner with a usp of 200 gal, and the mixture was washed to clean the pellet after centrifugation. The wet block is packed into a plastic or drum of a double-layered polyethylene bag.

1071-3452-PF.ptd Page 23 V. INSTRUCTIONS (16) 1282783 The N-(5-vaporized acid)-8-aminooctanoic acid is under vacuum in a vacuum (27 inches) dry. The dried cake was placed in a double-layered polyethylene bag, placed at 55 gallons, and the steel was placed inside the upper open drum and covered with a dehumidification bag. The dried product was 81 kg of N-(5-chloro!-sulphate)-8-aminooctanoic acid. j Example 2: Preparation of N-(5-chlorosalicylate)-8-aminooctanoic acid disodium salt I (disodium N^(5~chlorosaIicyl〇yi)_3-aminocaprylate)

A 2 liter five-piece dispenser, a thermocouple temperature reading, and a sodium hydroxide solution added to the stirred slurry of the keto-(5-chlorinated acid group)-8. The solids have all dissolved. This semi-permeable filter is hot filtered to remove the insoluble bottle of the inverting rotary concentrator. The RIS glass round bottom flask was placed at a temperature of 60 ° C with a bath temperature of a stirrer. To the flask was added 2 8 0 2. 3 -aminooctanoic acid and 4000 ml of water. When the mixture was heated from 660 g of water dissolved in 2000 ml of water to about 55 ° C, most of the bright solution was granulated with Whatman-filtered paper. The filter solution was poured into a laboratory with a large rotary concentrator at a pressure of 60 mm Hg. Moisture retention is only in the two sodium salt solution

1071-3452-PF.ptd Page 24

1282783 V. INSTRUCTIONS (17) Until solid substances appear in the concentrate bottle of the rotary concentrator. Destroy the vacuum and remove the bottle from the concentrator. The solid material is scraped from the bottle and loaded into the tray. The tray was then placed in a vacuum oven and the solid was dried at 60 ° C and maintained under vacuum for about 48 hours. The dried solids were ground through a laboratory with a grinder to a net of all solids that passed through a 35 m aperture. The sieved bismuth-(5-chlorinated acid)-8-aminooctanoic acid di-nano salt is placed in a tray and placed in a drying oven. Dry at 4 5 Torr and continue to maintain a full vacuum to obtain a powder product of 2 9 5 7 · 1 brother. The product was titrated with hydrochloric acid to obtain two inflection points, which consumed about 2 moles of hydrochloric acid. Carbon, hydrogen and nitrogen tiller: theoretically (corrected with 4.9 wt% water) carbon 47·89 % 'hydrogen: 5.37%, nitrogen 3.72%, sodium: 12.22%; actual value: Carbon 47.69 %, hydrogen: 5.23%, nitrogen 3.45%, sodium: 11.79%. Example 3: Preparation of N-(5-chlorolaurate)-8-aminonamate monosodium salt (monosod i um

N-(5-chlorosai icyl〇yl)-8-aininocaprylate A 22-litre five-piece Perez glass round-bottomed flask equipped with a stir state, thermocouple temperature reading, and a heating pack. To the flask was added 2 〇 99. g of N-(5-vaporized sulphate)-8-ammonium octanoic acid and 6 liters of water, and the mixture was dissolved. To the slurry was added a solution of 265 g of sodium hydroxide dissolved in 2 ml of methanol. The mixture is heated to about 8 Torr so that most of the :: the solution mixture is poured into a laboratory shrinking bottle. The rotary squeezing machine was concentrated at a pressure of 7 〇 璧 璧 璧 。 60 60 。 60 ° C. Moisture continues from: sodium salt;

1282783

Until the substance. The vacuum was removed and the solid material was scraped from the bottle. The solid was dried at 60 ° C and passed through a sieve of the Laboratory 罔 3 5 m e s h. After research 5, invention instructions (18) to the concentrated bottle in the rotary concentrator appeared solid impurities removed the concentrate bottle from the thickener. One of them is in the tray. The tray was then placed under vacuum and maintained under vacuum for approximately 48 hours. The dry and full grinder is ground until all solids can be sieved through the hole and sieved through N-(5-chlorinated acid)-8-aminooctanoic acid early salt 1 into a tray and placed in a drying oven. Drying under full vacuum gave 2 1 6 1 · 7 g of powder. The product was titrated with hydrochloric acid to obtain a reflex point, which consumed about 1 mole of hydrochloric acid. Analysis of carbon, hydrogen and nitrogen: theoretically (corrected with 〖·························································· Value: carbon 52·57 %, hydrogen: 5.66%, nitrogen 4.06%, sodium: 6.5 〇'%. 〆· Example 4 The case of applying the Cannes salt and the di-nano salt of 5-CNAC to the weeping macaque is as follows. A group of six monkeys were given a knee capsule containing disodium salt, and the second group of six clients were given a gel containing single sodium salt. Mg 5-CNAC (single or disodium salt) and 8 〇 () micrograms of squid #5 regulating hormone (sCT) to a hard gemidine gelatin. The above-mentioned weir monkeys were fasted overnight overnight before administration, and were kept on the r and awake during the experiment. The capsules were fed via a feeding tube and given 10 ml of water. Blood samples were taken at 15, 30 and 45 minutes and at 1, 1.5, 2, 3, 4, 5 and 6 hours after dosing. sCT concentration in plasma, radioimmunoassay

1282783

The test method is manic (^〇4 money 11110 &amp; 85 &amp; 7). In the two groups, 6/1^_隹* 丄. The eight blacksmiths averaged and plotted the results at each W point. In the average blood ride, the highest value of hormone concentration and its area under the curve (AUC) are listed in the first adjustment table 1 and table. Transmitting agent Transmitting agent Man sCT agent love Absorption 竣: hand-average blood aunt mother adjust the fresh lacquer degree ATJC — 1 (mg) (mg) (Pick / ml 士 标假假) 礼! i Γ 5-CANC 400 800 ^24±230 883 ~ disodium salt (94) 5-CANC 400 800 93.2 ± 133 16 broad sodium salt _ (54) Example 5: selection of appropriate starting materials, using Example 1 Method for the preparation of 彳Μ-(10-[2-hydroxybenzylphenanthryl]amino) decanoic acid u Example 6: Preparation of bismuthinoic acid-moxi-amino citrate disodium salt ( Disodium Ν - salicyloyl -10-ami nodeca n oate)

NaOH EtOhT

A 1 liter four-piece Perez glass round bottom flask equipped with a stirrer, reflux condenser, thermocouple temperature reading, and a heating pack. The burning ship

A 1071-3452-PF.otd Page 27 1282783 V. INSTRUCTIONS (2G) ~-- Washing with dry nitrogen, and the following reactions are carried out under atmospheric pressure. The flask was charged with 100 g of salicylic acid 10-amino hydrazine and 500 ml of absolute alcohol. The slurry is stirred and heated to about 4 Torr to dissolve the solids. An additional funnel was placed on the reactor to add 232 g of 11.2 wt% sodium hydroxide dissolved in absolute alcohol. The sodium hydroxide solution was added to the stirred mixture in the reactor over 15 minutes. ^ The 1 i stream condenser was removed from the reactor and charged with a distillation tap and receiver. The mixture in the reactor was distilled under atmospheric pressure and 395 g of j distillate was collected. The reactor mixture turned into a thick sludge in the steaming pin ‘ | the mixture was cooled to temperature. This mixture was poured into a frit of a sintered glass, and the solid matter was collected by vacuum filtration. The wet cake of ethanol was placed in a vacuum oven at | 4 5 ° C and dried to constant weight under full vacuum. The dried state of the state is about 124.6 grams. I titrate the product with hydrochloric acid to obtain two inflection points, which consume about 2 moles of hydrochloric acid. Carbon, hydrogen and nitrogen analysis: theoretically (with 0. 47wt% water for external positive) carbon 57. 15%, hydrogen: 7.37%, nitrogen 3. 51%, sodium · 11. 51%; actual value : carbon 57.30%, hydrogen: 7.32%, nitrogen 3.47%, sodium: 11.20%. Example 7: Preparation of ethanol solvate of disodium N-(5-chlorosalicyloyi-8-aminocaprylate)

1071-3452-PF.ptd Page 28 1282783

V. Description of invention (21) 〇

A 12-litre four-piece Perisian glass round bottom flask containing a drop, a thermocouple temperature dive, a reflux condenser, and a heating pack. The treatment was washed with dry nitrogen, and the following reactions were carried out under a dry atmosphere of atmospheric nitrogen. The flask was charged with 100 g of N-(5-chlorosulphate)-8-amino group; octanoic acid and 3000 ml of absolute alcohol. The mud 1 slurry was heated to about 55 C to obtain a semi-transparent solution. Add 2276 grams of π. 2wt% sodium hydroxide dissolved in absolute alcohol in the reactor. At this point, there is a slight exothermic reaction that causes the reaction temperature to rise to about 64t: at the same time, precipitation begins to occur. The reflux condenser was removed from the reactor and the reactor unit was taken care of. The mixture in the reactor was steamed for three hours to give a distillate of about 2,566. The reactor mixture became a thick slurry in steamed radium. The mixture in the flask was slowly lowered to room temperature. The loach mixture was vacuum filtered through a funnel of a sintered glass to collect 1 390 g of ethanol wet mass. The wet block is placed in a glass tray and placed in a vacuum for shooting. At 4 5 . (: Dry under full vacuum to 愠 weight. The product obtained by drying is about 1 094·7 g.'

1071-3452-PF.ptd Page 29 Meaning 1282783 V. INSTRUCTIONS (22) The product was titrated with hydrochloric acid to obtain two inflection points: about 2 moles of hydrochloric acid was consumed. Analysis of carbon, hydrogen and nitrogen: theoretically (corrected with 〇wt% water) carbon 50· 56 %, hydrogen: 5.99%, nitrogen 3.47 %, sodium: Η. 391 %; actual value: carbon 50.24 %, hydrogen: 5' 74%, nitrogen 3.50% (sodium is determined). Example 8: Preparation of N-(10-[2-hydroxybenzoguanidino]amino)oxime|monosodium 丨N-(10-[2-hydrobenzoyl]amino) decanoate! A 22-litre five-piece Perez glass round-bottomed flask equipped with a stirrer, thermocouple temperature reading, and a heating pack. To the flask was added 8 〇1·8 Ig of hydrazine-(10-[2-hydroxyphenothioni]amino) citric acid and 6 liters of water. A sodium hydroxide solution prepared by dissolving 4 g of 丨Q in 3 Torr (10) ml of water was added to the slurry. The mixture is heated to about 63 〇c to dissolve the solids of the large portion. The translucent mixture was poured into a laboratory-circulating large-scale spin-concentration machine. The water is continuously drawn from the monosodium salt solution until a solid substance appears in the concentrate bottle of the concentrator. Remove the vacuum and remove the &gt; dent bottle from the concentrator. The solid material is scraped from the bottle and loaded into the f tray. The tray was then placed in a vacuum oven and the solid was dried at 80 ° C for about 48 hours. The obtained solid matter was confirmed to be the single steel salt desired. The dry solid weight was 822.4 grams. ^ Titration of the product with hydrochloric acid to obtain a reflex point, about 1 mole of farmland owing rabbits, sputum and gas analysis: theoretically (corrected with 〇 · 5 4 9 wt% water) carbon 61. 65 %, gas: 7.37%, gas 4. 23%, nano: 6% %; | Actual value: carbon 61.72%, hydrogen: 7.38%, nitrogen 3.93%, sodium: 6 61

▲ %i 1282783 V. Description of invention (23) %. * Example 9 · Preparation of N-Liipyl-1 〇-amino phthalic acid di-nano salt ethanol solvate / hepatic acetyl sulphate capsule! N- sulphate 10-amino decanoic acid disodium salt (SNAD) Ethanol solvate was sieved through a 1-20 mesh screen. Scale &amp; 7·77 g of sieved SNAD disodium salt solvent I compound and placed in titanium. Scale 1 · 35 g of hepatic sulphate sodium salt usp (丨82 single | position / 3⁄4 g)' (purified from Scientific Protein Laboratories,

Inc. Waunakee, WI) was added to the SNAD disodium salt solvate in the mortar. The powder is evenly mixed by a spatula. The uniformly mixed powder was placed in a product! The pit V-blender shell (purchased from | Patterson-Kelley Co. of East Stroudsburg, PA) was mixed for about five minutes. The size of the hard gemidine transfer capsule (purchased from T o r p a c I n c. 〇 f I Fairfield, NJ) was manually filled with about 297 to 304 grams of SNAD disodium I salt solvate/hepatic acetylcholine powder. The average weight of the powder in the capsule was about 3004 mg' and the average total weight of the capsule (i.e., the weight of the capsule containing the powder) was about 387.25 g. Each capsule contains about 259.01 grams of SNAD disodium salt solvate and about 44.0 mg of hepatic cadaverine. Example 10: Preparation of SNAD monosodium salt/hepatic stagnation lozenge The preparation of S N A D Zhuo Na salt/hepatic cadherin suspect was as follows. s n A D is first sieved through 3 5mesh _ net. Scale 岀15〇·3g of SNAD, 27.33g of liver 〜~素纳盐 (purified from Scientific Protein Laboratories,

Inc. Waunakee, WI) 112· 43 grams of Avicel OPHlOl (available from FMA Corporation, lnc·, dE) v 6· 克 克 Ac — Di·-s〇 1〇, (by FMA

! $

1071-3452-PF.ptd Page 31 1282783 I. Inventions (24) |

Corpora t ion West) and 2.265 grams of talc (by Soct run ! . - i

Chemicals of New Brunswick, NJ) Powerful to 2 quarts of V-granted cultivator (purchased from Patterson-Ke1iey Co. of East I

Mix ξ I I Stroudsburg, PA) for about five minutes. The result of the final mixing was compressed into a sheet by a | ΕΚ-0 tablet compressor (Korsch America Inc., of Sumerset, | I NJU). The resulting sheet was shattered through a 20 mesh screen to produce jj ί ! 3. The talc of the 3.94 brother and the Ac_Di-Sol seen by d.25 add the above particles |

In I, mix in a 2 quart V-blender for about 5 minutes. 2.72 grams of hard I. magnesium stromate was added to the microparticles in the V_mixer and mixed for about 3 minutes. The resulting | | formulation is made into a tablet with a ΕΚ_0 tablet compressor. The average tablet weight is 320. 1 丨 83 mg. Boxing S embodiment ί].

I I | 4 cynomolgus macaque monkeys, 2 male and female, \ i average weight kilograms, two capsules prepared in Example 9. Each ί I monkey was dosed with 150 mg/kg of SNAD ethanol solvate di-sodium salt and approximately 30 mg/kg hepatic cadaverine. Dosage tests for administering capsules to animals are as follows. Animals were fasted before the administration of the drug (fasting within two hours after administration). The drinking water was given during the administration, and 4 〇 ml of the juice was given to the animals during the fasting period before and after the administration. The capsule is placed in a pill dispensing device which is a plastic tool having a trigger piston and a rubberized open tip to feed the capsule. The pill is administered to the animal's esophagus. Pressing the pill-administered plunger pushes the capsule from the rubber tip j end into the esophagus. The drug gun is then withdrawn. Keep the animal's mouth tightly closed, and give about 5 ml of reverse osmosis water from the mouth to guide the swallowing reaction \ &quot; ί

l〇71-3452-PF.ptd Page 32 1282783 V. Description of invention (25) should. At the same time, the animal's roaring Liu Qiu, J disease ^ to swallow the reaction, soil, '. TPC before the start of the order from 15, 9, 3, "b', two, moxibustion, 20, 30, 40 and 50 minutes and, 1 ϋ L ό 4 and 6 hours, * painful gentleman /... Blood sample taken from the vein: two veins (thigh, arm or hidden π ^ ^ 4- , , ... d pen liters. Blood sample and 0.31 wells about 0 · i 06M release salt solution隹...,... 古么ΛΑ W 一上&lt;Λ•不d into the official. Blood is added to the test tube I 3 2 24〇Ί Ί' Then the test tube is taken to the ice to wait for centrifugation. The liquid sample is too ^ 1 will CJ was centrifuged at 2~δ °C for 15 minutes at low temperature. The blood water was not suffocating, friend: 隹; ^ A y, . , 0^Λ In the analysis, stored in dry ice or frozen I ί UC 〇 丨 丨 I Plasma hemagglutinin concentration by anti-factor Xa assay 4 I (anti-Factor Xa assay CIIROMOSTATEO heparin anti-Xa jj assay, obtained from 〇rganon Teknika c〇rp〇rati〇n 〇f ― Juice (10),|

i nl) determination. The scars obtained from the animals are averaged at each time point. The maximum average reached 1 hr after I I administration was 154 ± 0.17 IU/ffiL. I! Comparative Example 11A | The procedure in Example 11 was repeated by substituting the SNAD monosodium salt bond prepared in Example 10 for the disodium salt of the SMAD ethanol solution. Average 4 ί

Two kilograms of monkeys were given two lozenges. The dose is about 150 mg / kg I :! SNAD (free acid equivalent) and 30 mg / kg of liver cadherin. The maximum mean plasma hemagglutinin reached 2 hours after administration of 0. 2 3 soil 〇 19 IU/mL. Example 12: Preparation of mono-sodium N-(8-[2-hydroxyoxy]amino)octanoate (Mono-Sodium N-(8-[2-hydroxy ben zoy'l] am i no )

1071-3452-PF.ptd Page 33 1282783 V. INSTRUCTIONS (26) j caprylate(SNAC)salt) ' j SNAC free acid (eg: n-(8 - [2-hydroxy methoxymethyl sulfhydryl]] Amino)|octanoic acid) was prepared by the method of Example 1 by selecting an appropriate material. j I Add 321 liters of ethanol to 5·5 % 曱 变性 and then add 300 gallons of |j reaction. The disturbance is set at 1 q q - 1 2 5 r ρ πι, and the reactor contents are lowered by a temperature of ~9. While stirring Φ, 1 〇 9 kg of dry free j·J acid was added. The reactor was heated to about 28 tons: and the temperature was maintained above 25 °C. 34 liters of j I pure water, USP mixed with 15.78 kg of sodium hydroxide, added to the stirred reaction for 15 minutes, maintaining the reactor temperature between 25-35 ° C. The mixture j was further stirred for 15 minutes. ^ In the second reactor, add 3 2 1 liter of Yi Yi, with 0 k 5 % 曱 变 ί · ^ k j j sex. The reactor was heated to 8.0 with a circulator. The solution i I in the first reactor was injected into the second reactor for 30 minutes 5 and maintained at a temperature above 25 °C. Stir | ^ I contents and add 418 liters of heptane. The mixture in the reactor was cooled to j j 10 °G, centrifuged, and rinsed with 60 liters of heptane. Collecting products and at j

The Stokes oven was dried at 82 with 26 Torr and vacuumed for about 65 hours (about 1 j weekend). 1 〇7.5 kg of a SNAC mono-salt salt (e.g., monosodium salt of (8-[2-hydroxo]phenylphenyl]amino)octanoic acid) is obtained. Example 13: Preparation of the disodium salt of SNAC The free acid of SNAC (e.g., Ν-(8-[2-hydroxyoxycarbamoyl]amino), octanoic acid) was prepared in the following manner. The sodium salt of SNAC i prepared in Example 12 was taken as j

One equivalent of concentrated hydrochloric acid is acidified and stirred in water. The solution was again filtered through vacuum and vacuum dried to give the free acid. I ί |

I Weigh 100 grams of SNAC free acid and add 2 liters of 4 rounds. I

1071-3452-PF.ptd Page 34 1282783 t V. INSTRUCTIONS (27) Bottles are filled with 500 ml of absolute alcohol. The temperature is set at about 40 ° C to dissolve the solid hydrazine into the liquid. The temperature rises to about 82. At 〇, 255·7 g of η· 2 /(w^w) sodium hydroxide ethanol solution was added to the solution through a sputum addition funnel over 15 minutes. At a temperature of 76-79 ° C, after about 1.5 hours, distilled W to 3 ^ 3 · 1 gram of ethanol. The mixture in the reactor was vacuum filtered through a coarse-pore funnel with nitrogen at about 2,000 ° C to afford a solid. Thus, the mass of f was washed with a filtrate and placed in an evaporation, and placed in a vacuum to dry 11 A at room temperature overnight. 90.5 g (68%) of a solid solid SNAC ethanol, a granulated product of a second steel salt. The melting point is greater than 2〇〇. 〇 (Instrument pole?) \HPtLCl check, showing 1〇G aera%. NMR showed the desired product. , 'Nitrogen and rat (^7H25N〇5Na2 * 〇·1265Η20) calculated values: carbon 54.94, nitrogen · 6.85, nitrogen 3. 77 ' sodium: 12 · 37 ; actual value: carbon 55 · 04, hydrogen: 6 · 56, ίΐ3·89, Na: 12 34. . A sodium salt monohydrate of SNAC was formed into an early hydrate by cooling the above ethanol solvate in C. After drying for 22·75 hours and then cooling to room temperature in an open space. The structure of the hydrate was confirmed by elemental analysis: C17H19N04Na2 · 0· 127Η20 Calculated value of carbon 53·〇1, hydrogen: 6.18, nitrogen 4.12, nano·15·53; Actual value: carbon 53 〇i ' Hydrogen: 61〇, nitrogen 3.88 ^ ^ · 13.08 〇iT〇lHNMR (300MHz, DMSO~d6): dl2.35(lH, s), 7.55 (1 H, dd), 6.8 (1 H, dt), 6·25 ( 1Η, dd), 6.00C1H, dt), 3.2(2H? q), 1.9(2H, t), ' 1·45(4Η,bc〇,ι·25(6Η,bm). Melting point greater than 25(pc (Instrument limit!) I Example 14: Preparation of the monosodium salt of SNAD,

1071-3452-PF.ptd Page 35 1282783 V. INSTRUCTIONS (28) The acic acid of SNAD can be prepared by selecting an appropriate starting material, which is inexpensive. “ ^ Inject 206 liters of SNAD containing 33·87 kg of 〇·5 % 曱 benzene into the reactor. Stir for one hour and then pass through a pressure filter. ^I 么 的 石 石 ( ( (by Celite Corporation oi Lomp〇c, I got) added to the reactor. The mixture in the reactor was pressurized, and the sonic solution was left in the separator. The reactor was washed with five gallons of deionized water. Effluent with sodium hydroxide solution (4 · 5 kg of sodium hydroxide and 丨 2 Gongfeng,

I water) was introduced into the reactor. The mixture in the reactor was disturbed for 30 minutes, and the J j temperature was removed by vacuum to remove 3 gallons of solvent. Mix the parts in the reactor to 60 C, then pour into two 1 〇〇 gallon tanks on average, and add the same heptane to each of the two tanks and stir quickly for 2 hours. The solution was centrifuged | 15 g after the g-burning, spray-dried, and dried in a 45 t: oven in the right '" column for 24 hours. The crucible was passed through the Fitzmill grinding state (by Fitzpatrick Company). Oi Elmhurst, IL obtained) Grinding. 32 kg of smear powder SfD monosodium salt (melting point 190-1 92 t, HPLC analysis carbon: 99j % color · d7). The titration results show that it is about 96 % SNAD single Sodium gg 盥 4 % SNAD disodium salt., two" ▲ Example 15: Preparation of SNAD disodium salt SMDdClG - [2 - hydroxy alkanoyl] amino) decanoic acid) Free S owage can be selected by Y A suitable starting material was prepared according to the procedure of Example i.

1071-3452-PF.ptd Page 36, add 100 grams of SNAD free acid to a four-neck round bottom flask, and add 500 liters of absolute alcohol. The temperature was set at 40 ° C to dissolve the solid in the liquid to give a pale orange solution. 23 2. 5 g of 11 · 2 (, w / w) % hydrogen 1282783 V. Description of the invention (29) '^ - The sodium oxide alcohol solution was slowly added to the flask through the additional column for 5 minutes, and the temperature rose to about 8 2 C. After 3 hours, the temperature is between 7 5 and 7 9 . . 1 Distillate 39 7·8 g of ethanol. The mixture in the reactor was slowly under nitrogen overnight... to room temperature. The resulting slurry was vacuum filtered through a coarse aperture funnel to give a solid which was washed with filtrate. The damp filter cake was placed in a steaming dish and placed in an oven at 50 ° C and dried under vacuum overnight. 124.55 grams of SNAD monosodium salt (96%) gave a pale pink ethanol solvate solid. The melting point is below 200 °C (instrument limit). The HPLC trace shows a 1% peak area. NMR was not the product. The calculated value of CNiI is carbon 57·42, hydrogen·· 7.35 'nitrogen 3.52, sodium: 11.57; actual value: carbon 57.37, hydrogen: 7.35, nitrogen 3.41, sodium: 11 · 63. The disodium salt monohydrate of SNAD was prepared by solvating the ethanol at full vacuum for more than 80 t for 19 hours and then reducing the solution in air to a temperature to form a monohydrate. The hydrate structure was confirmed by elemental analysis. C17H23N04Na2 · H20 calculated value of carbon 55·28, hydrogen: 6.82, nitrogen 3.79, sodium: 12·45; actual value: carbon 56·〇3, hydrogen: 6·67 , nitrogen 3.67, nano: 12·20. And 1H NMR (300MHz, DMSO-d6): dl2.35 (lH, s), 7·6 (1Η, dd), 6·8 (1Η, dt), 6·25 (1Η, dd), 6·00 ( Η, dt), 3·2 (2Η, q), 2·0 (2Η, t), 1·45 (4Η, bt), 1·25 (10Η, bm). The melting point is greater than 25 (TC (instrument limit). Example 1 6: Oral delivery agent for hepatic acetylcholine contains oral sodium of heparin sodium USP and the single sodium salt or di-nano salt of the delivery agent component SNAC (p 〇 The administration solution is prepared in water, and the delivery agent component is mixed with hepatic calcin (166 IU/mg) in a dry powder form by vortex stirring.

1071-3452-FF-pid Page 37 1282783 V. INSTRUCTIONS (30) The compound is dissolved in water and at 37. (: Mix with vortex and sonic wave to form permeation / 1 / valley solution. P [{ value is not adjusted. The final volume is adjusted to 1Q Q ml. The final delivery agent component dose 'hepatic cadherin dose and dose volume is listed in The following is the first! Table 2. ϊ ! The general dosage and sampling schedule are as follows. The average weight of 2 75 5 to 3 50 | grams of Sprague-Daw ley male rats after fasting for 24 hours, before and after administration Keta was anesthetized by intramuscular injection of hydrochloric acid (ke t am i hydroch 1 or i de, about 88 mg / kg), and was re-injected as needed to maintain anesthesia. The number of single doses was 1Q mice. The 1 ml syringe of the liquid tube is connected to the I 1 1 cm Roche 8 French tube. The syringe is fed into the j solution through the fistula catheter stage, and then the catheter is wiped dry. The catheter is placed in the esophagus and left in the centimeters. The incisors are protruded. The dosing solution is injected with a citrate-containing blood sample by pushing the plunger of the syringe, and is obtained by cardiac puncture at 〇·25, 〇·5, 1.0, and 1-5 hours after administration. Absorption of sputum The ial thromop last in time, ΑΡΊΤ) method demonstrates an increase in the clotting time (according to jB· Henry, Clinical Diagnosis and Management by Laboratory Methods, Philadelphia, PA, WB· Sauders, 1979·) 〇 Previous studies showed no reference value of approximately 20 Seconds, and the average value of each group of animals at each sampling time and the T value of the LIda AP (in seconds) are listed in the second fungus. Anti-factor Xa CHROMOSTRATE® Heparin anti -Xa assay ^ Organon Teknika Corporation of Derham, NC.) Measuring liver, condensate

1071-3452-PF.ptd 38 I 1282783

The concentration in the mortar demonstrates an increase in hepatic calcin. The baseline value is approximately ου I/mL. The plasma hepatic concentration of the liver at each sampling time of each group of animals was averaged and greened. The mean plasma hemagglutinin concentration wave therapy values are listed in Table 2. The second component of the drug administration (ml / kg) component agent Man (mg / kg) liver sputum 壷 (mg / kg) average peak APTT (seconds) Shi SE · average wave peak Xa factor soil SE SNAC single Sodium salt 300 100 24 soil 728.5 2.597+0.13 SNAC disodium salt 3 300 --- 100 1 30〇±〇1 2.8110.17 i Example 1 7: Low molecular weight hepatic clotting agent oral delivery agent (Low

Molecular Weight Heparin, LMWH) An oral administration (P〇) composition containing a low molecular weight hepatic condensate (LMWH) and a monosaccharide sodium or disodium salt of the delivery agent component SNAD is prepared in water. Transfer agent! Ingredients with 1^!丨?11 (?31'113?81'丨!1,9111[/1^, average molecular weight 5000, ϊI from Opaocrin of Modena, Italy) in the form of a dry powder vortex Stir and mix. The dry powder mixture was dissolved in water and mixed with a vortex and a sonic wave at 37 ° C to form a clear solution. The PH value is not adjusted. The final volume was adjusted to 10.0 ml. The final delivery agent dose, LMWH dose and dose | volume are listed in Table 3 below. The administration was the same as in the above Example 13. Blood samples containing citrate were obtained by cardiac puncture at 05, 1·〇, 2·0, 3·〇 and 4. 0 hours after administration. Measurement of the concentration of hepatic statin in plasma by anti-Factor Xa CHROMOSTRATE® Heparin anti-Xa I assay, Organon Teknika Corporation of Derham, NC. I demonstrates an increase in hepatic pacingin. first

1071-3452-PF.pid Page 39 1282783

The baseline value for 刖 determination is approximately 0 UI /mL. The concentrations of f-containing hemagglutinin at each sampling time for each group of animals were averaged and plotted. The mean peak value of plasma hepatic condensate concentration is listed in Table 3. Volume 3 ingredient dosage volume (ml/kg) Component agent Man (grams/kg) LMWH dose (mg/kg) Average peak liver cadmium concentration (IU/mL) Soil SE SNAD Single sodium salt 3 300 3000 0· 8 belly 0.17 SNAD disodium salt 3 300 3000 1.21±0.15 Example 18: Preparation of N-(5-chlorolauric acid)-8-aminooctanoic acid | (5-CNAC) | ! Under nitrogen 5-chlorolauric acid (280 g, 1.6 mol) and acetonitrile 6 70 ml were placed in a 5 liter 4-neck round bottom flask and stirred. 161· 3 gram mouth bite (2·0 mol) is slowly added to the above mixture within 25 minutes. The reaction vessel was placed in an ice water bath and ethyl chi orof or mate was added in a medium ratio at one hour. When the addition was complete, the ice water bath was removed and the reaction mixture was allowed to warm to room temperature. The reaction mixture was stirred at room temperature for an additional hour, after which the reaction vessel was steamed at atmospheric pressure. When the upper end temperature was 78 ° C, 257.2 g of distillate was produced. 5 ml of deionized water was added to the remaining mixture in the flask, and the resulting slurry was vacuum filtered. The filtered cake was washed with 200 mL of deionized water and dried overnight at room temperature under vacuum. After drying, 31 3 · 6 g of 6-chloro carsalam (97·3%) were obtained. The other batch was also prepared in the same manner as 44.5 g of 6-chloro-2H-1,3-benzoquinone-2,4(3H)-diindole J.

1071-3452-FF-ptd Page 40 1282783 V. INSTRUCTIONS (33) 1.8 mol of 194.0 g of sodium carbonate was added to 6-gas-2H-1,3 containing ^23·1 g (16 mol). A 5 liter, 4-neck round bottom blister of phenylmorphine-2,4(3H)-dione and 970 ml of dimethyl]ethenolamine. An equal ratio of 4 5 9 grams of |ethyl-8-bromide ruthenium (1.8 moles) in the above mixture of the mixture was reacted | the pressure in each was reduced to 550 mm Hg, i The heating of the mixture was started. j The reaction temperature was maintained at 70 ° C for about 5 hours, after which the vacuum was released and the addition of temperature was stopped. The mixture was cooled overnight. The reaction mixture was then vacuum filtered, filtered, and the cake was washed with EtOAc. 525 liters of deionized water was added to the stirred filtrate to form a white solid precipitate. The vessel of reaction j was placed in an ice water bath and the slurry was cooled to 5 liters. After stirring for about 1 b minutes, the solid was separated by vacuum filtration, and the solid was washed with 300 ml of ethanol and then washed with 4 ml of heptane. After drying in vacuum at medium to warm overnight, a sulphuric acid salt of 8-8-(6-gastriene, 3-brown--2,4(3H)-) octanoate was obtained. Gas 1 Add 1 ♦ 7 moles of 64 μg of ethyl 8-(6-nitro-2H-1,3- alum 2,4(3H)-di-iso)octanoate with 3200 ml of ethanol to 22 liters In a 5-neck round bottom flask. In a separate 5 liter flask, 288. 5 grams of 7.2, sodium hydroxide was dissolved in 3850 milliliters of deionized water. The sodium hydroxide solution was added to a 22 liter flask and the temperature was raised to 4 Torr. The above mixture was heated and all solids were dissolved at a temperature of 5 °C. At this time, the temperature was maintained at 50 ° c i · b hours. The reacted flask was then subjected to distillation. When the steam temperature is 55 °C (10 mmHg), the concentration of 22 〇〇 ml of the smash is collected. ^ This view is placed in the ice water. Slowly add concentrated hydrochloric acid 7 5 2 4 in 45 minutes. Rise. In the added mash, the mixture will become rich, because

1071-3452-PF.ptd Page 41 1282783 I —---- ---------1 ! V. Description of the invention (34) 1 I Add 4 liters of deionized water to help stir. The mixture was filtered through a vacuum, filtered, and the cake was washed with 3 liters of deionized water. After drying at room temperature, 456. 7 g of N-(5-chlorosulphate)-8-amino-1 |octanoic acid (83.5%) was obtained. ί | Example 19: Calcium calcium regulating hormone (sCT) and 5-CNAC sodium salt were cooled. i | 5-CNAC sodium salt damage | I The purity percentage of 5-CNAC is determined by the following method. 0. 9964 g | I 5-CNAC free acid dissolved in 40 ml of sterol. After dissolution, 2 milliliters of | liter of distilled water was added. The above methanol solution was titrated with a sodium hydroxide solution of 〇 · 3 3 N via a computer controlled drop _ |Hami1 ton automatic burette (from Hamilton 〇f | I Reno, NV). The pH of the solution was monitored with a glass electrode (computer controlled ! I ί j Orion model 525A pH meter 5 from VWR Scientific οί | I South P1 a i n; f i e 1 d, N J ). The solution was mixed with a magnetic j 丨 agitator. 1

The titration volume reaching the inflection point of the second dH was 18.80 ml. The inflection point ph i 1 · | 3 of the second from the positive to the negative position of the I * p | pH pattern. For the calculation of free acid purity, see the following formula: j Purity (%) = (100x (ml of titration) x equivalent concentration x molecular j) / (100 X equivalent X sample weight) . 5 equivalents of sodium hydroxide Equivalent concentration; molecular weight is j | -CNAC free acid molecular weight; equivalent is free acid equivalent (in this case | | 2, because it is 2 bases); and the sample weight is the weight of the titrated free acid sample | r $ i amount. !

1071-3452-PF.ptd Page 42 1282783 V. INSTRUCTIONS (35) Purity is 9 7 · 〇 %. 'By 9 * 3 4 5 8 grams of 5 - C N A C powder eight. According to the formula, the molar ratio of sodium sulphide to 3 3 N to free acid is 1.6. Sodium hydroxide volume (ml) = (free acid weight X purity (by χ i 〇 〇 X 6 ) / ( 3 1 3. 78 X 1 00 X equivalent concentration) &quot;&quot;

Wherein, the free acid weight is a free acid sample according to the structural formula; the purity | (%) is the purity percentage of 5 - CMAC; the equivalent concentration is the concentration of the hydrogenation sodium; and the volume of the sodium hydroxide is sodium hydroxide The total need for I3 I 5-CNAC is mixed with 1 53·3 ml of 0·33N sodium hydroxide in a Peres glass bottle | The resulting slurry was heated to a temperature of β〇~80 °C in a steam bath. The ρ Η value of the solution was 8.1. Preparation of j sCT/5-CNAC sodium salt solution | 5 —Aqueous solution of CNAC sodium salt via a Corning filter (from VWPv Scientific Pr〇duct) with sterilized 〇 45mw acetate, low protein binding filter, S. Piainiield, NJ) Filtration. 5。 The pH of the solution is about 8.3. The dried salmon squid calcium-regulating hormone (SCT) stored in -70 t: is returned to the temperature. Weigh 1 8 · 6 9 2 s sCT dissolved in 1 〇 well 〇 · 1 μ, about pH 5 in a single sodium silicate buffer solution, and gently mix. SC1 ling solution is added to the 5-CiNAC sodium salt solution and gently stirred, taking care to avoid blistering or vortexing. °

!

*, 'Tutong, ^ regulating hormone (sCT) and 5-CNAC sodium transfer

107!-3452-?F.ptd第43页

1282783 V. INSTRUCTIONS (36) Approximately 260 ml of (sCT) and 5-CNAC sodium salt solution was poured into a stainless steel tray of 30 cm x 18 cm, which was about 4.8 cm higher after injection. Insert four clean and dry thermocouple probes into the solution at approximately the middle of the solution. The probe is clipped to the side of the tray and the tray is fed to the shelf of the pre-cooled freeze dryer. In the fourth table, there is a gel permati0n chromatography method (GPC2) listed in the flow table 4: cold; east drying process step temperature pressure setting (亳托耳) _ 1 丨45X: 120 2 | -30 °C 1 300 ISO 3 j -20C Γ 200~~ 200 200 L~4~~1 ~~40^~ 360 ^ I 〇C [ 200 ^ 720 6 | 10 °〇100 ! 540 | —7 ~8~~ 20° 〇卜坑~ 100 ------- 100 -36〇_j 180 | . During lyophilization, the pressure is changed from 305 to 45 millitons. When the freeze-drying process is stopped, the system stops and vents the vacuum. , 丨 心 心 由 洽 洽 j j j j j j j j j j j j j j j j j j j j j j j j j j j j j &quot; Fine from the above freeze-drying process, a composition having a water content of about 3% is obtained. The powder was hand-filled into hard gillidine capsules (size 0EL/CS) (available from Capsugei, a divisi〇n 〇f Warner “heart to c〇 〇ι:

Greenwood’ SC). The filled capsules and powder are stored in a closed container containing a desiccant. 1282783

Example 20: Preparation of unseduled (sCT) 舆5-CNAC sodium salt | 56.81 ml (61.47 g, 〇6〇26 mol) of glacial acetic acid, 1 〇〇 00 g (L5794 mol) of 5 The chlorosalicylic acid and 2 liters of hydrazine were added to a 500 liter 3-head round bottom flask with a magnetic stirrer, a thermometer and a condenser with a Dean_stark elbow. The flask was heated to reflux, and the reaction solution turned yellow from clear to about 100 °C. Most of the volatile organic components (eg, bismuth and acetic acid) were dehydrated by Dean-Stark elbow (1 35-1 46 C and the distillation was continued for an hour, the temperature of the flask slowly rose to 丨g 〇c Only a small amount of solvent was distilled off. A total of about 25 ml of the solvent was collected. The residue in the flask was cooled to below 丨〇〇艽 and then dioxane was added (dioxane) 222 222·85 ml 2N sodium hydroxide (〇·4457 mol) with 7 (K 96 g of 8-aminooctanoic acid (0.44457 mol) added with 〇·5794 mol oligo (cyclohexanoic acid) (〇14〇( 5-(:}11〇1*0 "14714"1(1)) in dioxane solution ~. The mixture was heated to 90 ° for 5 hours, then allowed to stand overnight. Reheated back the next morning ( Monitoring by HPLC was carried out during reheating &amp; when the reaction was stopped. The reaction mixture was cooled to 4 Torr. The dioxane was removed under vacuum. The residue was dissolved in 2N sodium hydroxide and then acidified. The material was extracted with ethyl acetate (2 χ 2 (10) mL) to remove excess dioxane. The sodium is dried and colloided under vacuum. The easily filtered solids are collected via filtration. The remaining material is dissolved in other sodium hydroxide. The pH is adjusted to 43 to separate the product from the starting material. Under ρΗ4·3, After passing through the filtration, the solid is recrystallized in a mixture of ethanol and water of J: j. Initially, all the insoluble matter is filtered out by heat. All 1282783 V. Inventions (38) ---~- -- The solids are collected and recrystallized from a mixture of ethanol and water.

| 5 2 · 0 6 g of white free acid solid. I. The sodium salt solution was prepared according to the method described in Example 19. When using 0·5038 g of milliliters of sodium hydroxide, the purity ratio was calculated to be 100%. The sodium salt solution was prepared according to the above method j · | with 2 50 ml of 〇 2 Ν sodium hydroxide solution and ‘458 5 gram of 5-CNAC. The solution was then filtered through a filter of 〇·45//πι. | j 灵知例21 · Mouse sCT/5 - CNAC sodium salt oral delivery agent to transfer j! Average weight to 250 grams of Sprague-Dawiey male rats fasting 24 small j! After the time 'i5 minutes before dosing Ethamamine (about 44 mg / kg) ^ | Anesthesia with hibernation (chl〇rpromazine, about mg / kg). The mice were administered in one of the following conditions: i | &gt; Ua) Oral administration, containing the freeze-dried powder i3 | j mg prepared in Example 19, one capsule with 5 ml of water; j I ( 4b) Oral administration, containing U mg/kg of the recombinant aqueous solution of the cold-dried powder prepared in Example 19; (4c) orally, fresh un-freeze-dried 5 prepared in Example 2 containing 1 mg/kg -CNAC sodium salt and sCT aqueous solution; or j (4d) subcutaneous administration, 5 mg/kg SCT. j (4a), (4b) and (4c) each contained 50 mg/kg of 5-CNAC sodium salt and 100 mg/kg of sCT. The dose of (4a) is about the amount, because the dose in the administered capsule is calculated as the average of 250 grams, and in fact the body weight varies. The experiment of the capsules mentioned later is also the same. ' i

1282783 V. INSTRUCTION DESCRIPTION (39) The recombinant solution in (4b) is prepared by dissolving 150 mg of the freeze-dried powder prepared in Example 19 in 3 ml of water and mixing the solution. The amount is 1.0 ml. /mg. The fresh solution of I (4c) was prepared by dissolving 150 mg of unsalted I-dried 5-CNAC sodium salt in Example 20 in 3 ml of water with 丨50 ml of sCT-based solution (200 ml/ml). It was prepared in 1 Μ phosphate buffer and the pH was adjusted to 4 with HC1 and I NaOH. The final concentration of fresh solution is 50 mg / ml

I 5-CNAC sodium salt and 100 mg/m well sCT, the dosage is LQ liter / male I i kg c j Subcutaneous injection is dissolved in 1 ml of water with 2 mg of sCT. Then take 1 ml of this solution and add it to 99 5 ml of water. 5毫升/j公斤。 The dosage of this solution is 0. 5 ml / j kg. I Blood sample The tail artery is sampled on time. The sCT concentration in the serum was determined by | EIA kit (Kit #EIAS-60 0 3, Peninsula I Laboratories, Inc., San Carios, CA), and the assay conditions were modified as follows: 50 ml of the peptide antibody was shaken under dark Incubate for 2 hours, clean the plate, add serum and biotin-chemical peptide, and inject it in 4 ml of buffer, and shake overnight in the dark. The results are shown in Table 5. 'i

1071-3452-PF.ptd Page 47 1282783 j V. Description of invention (40) Table 5: sCTAS-CNAC sodium salt in mice for oral administration 1 1 5-CNAC sodium salt f! (mg/kg SCT agent man (mg/kg) average peak serum sCT soil SD (pick/ml) (four) knee capsule 50&quot; 100* 1449±2307 (4b) recombinant solution 50 100 257±326 ~ (4c) unfrozen Dry sputum solution 50 100 134±165 (4d) subcutaneously to bran! 5 965±848 1 : Due to the difference in burdock weight, it is also about U. I Example 22: sCT/5-CNAC sodium salt oral delivery agent for mice, I According to the method described in Example 21, the mice were given the following two drugs: (5a) Oral administration, containing freeze-dried powder 1 3 mg capsules ~ 1 with 1 ml water; s I (5b) orally, containing 6.5 mg capsules of lyophilized powder - fed with ί ml of water; &amp; (5c) Oral administration, containing lyophilized powder 3. 25 mg of capsules are fed with 1 ml of water; or (5d) subcutaneously, 5 mg/kg of sCT. The dose and the amount of the above-mentioned delivery agent and sCT are shown in Table 6. Table 6: mouse sCT/5-CNAC sodium salt oral delivery agent dosage form 5-CNAC sodium salt agent worry (亳 / kg) sCT agent f ^ (mg / kg) hand average peak Jinqing sCT soil SD ( Pique / soaring) (5a) cane pouch 50 100^: ; 丨 379 456 (5b) wins 25&quot;50&quot; ! : Ιβ8±241 (5c) capsule 12.5* 25* | 0 (5d) subcutaneously Medicine — 5 Ί 273±320 : The difference in the weight of the animal, so it is about Man C, 1282783 V. Invention description (41) |

Example 2 3: Preparation of N-(5-chlorosulphate) 4-amino-butyric acid I; (N-(5-chi〇rosalicyl〇yi)-4aminobutyricacid) [30 g of sodium carbonate (〇·2) 8 3 5 moles) contains 50 grams of ethyl 6-chloro | -2H-1,3- alum 2, 4(3H)-dioxime (〇· 2 532 Mo, Example 18! Preparation ) 50 ml 3 neck round bottom flask with 75 ml of dimercaptoacetamide |

Medium and stir it. A further proportion of 45.83 g of decyl-4-bromobutyric acid I-^ (methyl-4-bromobutyrate; 0·2532 molar) was added and heating was started. | j The reaction temperature was maintained at 70 °C and heated overnight. When the heating was stopped, the mixed material was allowed to cool to room temperature. j j The above mixture was vacuum filtered, and the filtered cake was washed with ethanol. This cake was separated from the filtrate by HPLC to determine its composition. The products of Datun are | washed into the filtrate, but there are still a few left in the cake. The product in the cake is then separated to increase the yield. The cake was first washed with a large amount of water, and i group 1 | I was rinsed with ethyl acetate. The rinsing liquid was separately collected, and the ethyl acetate layer was washed twice with water | I ? and then washed once with concentrated brine. Dry over sodium sulfate, separate | and concentrate in vacuo to give more solid (solid B). A solid was precipitated after the addition of water to the previous filtrate j. The above solid (solid A) was isolated. The solid and B were combined and poured into a round bottom flask, n sodium hydroxide was added, and the mixture was stirred with heating. The reaction was determined by HPLC sampling to determine the degree of completion of the reaction. The reaction was cooled to 25 |1 〇 ° C., stirred overnight and concentrated in vacuo to remove excess ethanol. The reaction vessel was placed in an ice water bath and the mud was acidified therein. The solid material was obtained by vacuum filtration. The solid cake was washed with water and dried and then taken to NMR. The solid was separated and placed in an Er 1 e n m e v e r flask for recrystallization. Solid i

I analysis. j 1071-3452-PF.otd Page 49 1282783 V. INSTRUCTIONS (42) Recrystallization is carried out by crude alcohol/water. The crystallized solid was washed into a Buchner funnel. More solids in the eluate precipitated and separated. The solid separated after recrystallization forms a methyl ester. All solids were combined, and after 2N Na〇H was added, I was heated to reflux to give the free acid. When the ester disappeared (measured by 甴Ηριχ), the | mixture was acidified to a solution of about 4.7 to form a solid. I will separate the solids by filtration, and lick all the solids to recrystallize with 1:10 hydrazines and water. The white precipitated solid was separated and dried overnight to give &lt;RTI ID=0.0&gt;&gt;&gt;&gt; [,, ‘Measure 疋’ After the transition, the cakes are washed with excess ethanol for the first time to avoid product residue in the cake. Thereafter, the filtrate was stirred with 2N Na〇H, heated to 25 t and concentrated in vacuo to remove excess ethanol. = Ice water, the mud is acidified to pH 4.7. The cake was then vacuum filtered to remove the cake and washed with water. This solid was further separated and recrystallized. Example 24: sCT and Ν-(5-chlorosulphate) 4-aminobutyric acid sodium _ freeze-drying According to the method of Example 19, sCT/N-(5-chlorosulphate) 4 was produced. The lyophilized powder of sodium amide is loaded into a capsule. 10.528 g of N-(5-chlorosulphate) 4-aminobutyric acid prepared in Example 2 3 was dissolved in 150 ml of water' and 4.72 ml of ION NaOH was added. 21·0566 mg of SCT was dissolved in 10 ml of buffered phosphoric acid solution. The sCT/buffered phosphoric acid solution was added to the transfer agent solution, and water was added to make a total volume of 250 ml. Example 25: sCT/N-(5-chlorosulphate) 4-aminobutyrate sodium salt in mice, oral delivery agent administration,

1282783 I -- 丨丨 Surface | V. Description of the invention (45) t | Sex) Pour into the first reactor and warm to 45 °C. And 'filtered cake! Wash with warm ethanol and pour into the second reactor. Stir with a stirrer | 1 The contents of the 2nd reactor. The contents of the reactor were cooled to 2 9 | °C. 1 20 liters of deionized water is slowly added to the second reactor. Cool down its contents to 8t:. The intermediate product was separated from the solution and continued for 9.5 hours. The final slurry is centrifuged. Pour 70 liters of ethanol in the reaction 1 I, and cool to 8 ° C and wash the cake after centrifugation. The wet cake was placed in a double-layered polyethylene bag and stored in a container with a paper liner.得 | to i23.5 kg of ethyl 8-(6-gas-2H-1,3-stup π well-2,4(3H)-dione) octanoate. j I In a 200-gallon glass-lined reactor, add 400 liters of pure j water, USP and 45.4 kg of NaOH pellets, and the stirrer is set to | 100-125 rpm = ί23·5 kg Ethyl 8-(6-gas-2H-ί,3-benzoquinone-2, I 4(3Η)_dioxin) per acid salt cake was poured into the reactor and the injection was closed. The condenser is filled with condensed water. The valve above the reactor is set to atmospheric distillation. The mixture in the reactor was heated to 98 t: and monitored by HPLC. At about 4 Torr, the reactor was refluxed at 68 ° C, and after about 3 hours of removal of the ethanol by distillation, the reactor was raised to 98 °C. After initial separation by HPLC, the starting material disappeared after 4 hours. The contents of the reactor were cooled to 27 °C. 150 liters of pure water and USP were injected into another gallon of glass-lined reactor with the stirrer set at 100-125 rpm. 1 04 of concentrated concentrated hydrochloric acid (1 2 M) was injected into the reactor and cooled to 24 °C. The mixture of the saponification reaction was slowly poured into a 200-gallon glass-lined reactor over 5 hours. In the case of carbon dioxide production, this material is placed in two 2 gallon glass with 45 liters and 45 liters.

107!-3452-PF.ptd Page 53 1282783

Five 'Inventions' (46) Lining in the reactor. The product precipitated out of solution. The reaction mixture is adjusted to pH 2. 〇-4· 0 j between 50% NaOH (2 kg NaOH dissolved in 2 liters of water). Cool down to 9 - 1 5 C. The intermediate product is turned from the solution to j after about g hours. The mud in the reaction is centrifuged to separate the intermediate product.升 Rope water and USP are poured into a 200-gallon lining reactor^ to clean the centrifuged cake. The wet cake is placed in a double-layered polyethylene bag/j and stored in a container with a paper liner. 1 (5-chlorinated acid group)-g group, j octanoic acid was dried at 68 ° C for 38 hours under a vacuum of 27 Torr. The dry =| product is placed in a double-layered polyethylene bag and stored in a 55-gallon unlined enamel! The open container 5 is filled with a dry bag. After drying, it yielded 8 μg of W ^ 丨, (5 - gasified salicylic acid 彡-8-aminooctanoic acid. j Example 27: sCT/5-CNAC sodium salt was freeze-dried as a tablet poem I The 5-CNAC prepared in Example 26 was taken, and the lyophilized powder was prepared by using the method of Example 19. The solution was prepared by dissolving 42 g of NaOH in 2000 ml of water to prepare a sodium hydroxide solution. The mixture was stirred at room temperature and vacuum filtered through a 0.45 #丨m filter. The pH of the solution containing 5-CNAC was about 8.6. Using | 200 mg of sCT. j Example 28: Preparation of sCT/5-CNAC sodium salt Lozenge j The method of preparing the enamel tablet from the freeze-dried powder of Example 27 is as follows. Using Carver press (Model C) (from Carver οί |

Wabash, Indiana) is used to compress tablets. The steel mold of the circumference is the diameter | | 〇·245吋. The upper punch is a flat surface with a beveled edge and a diameter j | 0· 245吋. The lower punch is a flat surface, counting, edge bevel and 1

1071-3452-PF.ptd Page 54 1282783 V. INSTRUCTIONS (47) I Diameter 〇 · 245吋. Pushing above and below and removing the pressure pushed by the X side can be measured. The formula for direct compression is shown in Table 8. \第表表材料充充/锐!亳克/300 spindle batch SCT/5-CNAC sodium salt freeze-dried powder 100.2 j 30060.0 ( ί I AC-DI-SOL® 2.004 1 601.2 magnesium stearate 0.511 1 153.3 CAB-O-SIL0 0.205 Ί δΐΤ5 j Total Weight (亳先) 102.92 ί 30876.0 AC-DI-SOL® ^ croscraemellose sodium(N?, PH.Eur., JPE) 5 from FMC Corporation, Pharmaceutical Division, of Philadelphia, PA .CAB-O-SIL®: Smoked cockroach, available from Cabot Tuscola, IL.

I AC-Di-SOL® and CAB-0-SIL® are weighed and placed in a mixing bottle. The I bottle was placed on the operating arm of the slow release device and set to 25 rpm. This device is placed for 5 minutes for mixing. The freeze-dried powder of sCT/5-CNAC sodium salt was added to the mixing test of A C - D I - S 0 L ® and C A B - 0 - S I L ® in equal proportions, and the mixing was carried out at intervals of 2 minutes. Magnesium stearate was added to the above mixture and mixed for 5 minutes. About 103 mg of the above powder was placed in a steel mold containing a lower punch. The powder is pressed into the steel mold with an upper punch. The upper punch is inserted, and the fitting of the stamped steel die under pressure just dies and forms a squeeze. The upper punch then pushes the tablet out of the steel mold. Example 29: sCT/5-CNAC sodium salt oral delivery agent tablet for rats. Tablets prepared in Example 28 were ground into powder and manually filled into glue.

1071-3452-PF.ptd Page 55 1282783 V. INSTRUCTIONS (48) In the sac, 13 mg/capsule. In the case of Example 27, the frozen product was not compressed into a suspect! The dry powder was also filled in 13 mg/capsule. Each capsule is matched with 1 ml of j

Water is administered. I follow the steps of the embodiment, but make the same standard as the EIA suite. Rats are administered orally, each capsule is combined with 1 ml of water, and |

The dose and administration results of sCT/5-CNAC sodium salt are shown in Table 9. I Table 9: SC175-CNAC sodium salt oral delivery tablets for rats. 堃J sCT/5-CNAC sodium salt 箜I (mg/kg)]__ sCT agent 箜丨 hand peak f mg/kg) 1 soil · Qing SCT Shi SD 1 (pick / ml) (12a) capsule containing powder tablets | 50 * 100 * i 198 ± 132 I (12b) containing non-tablet end of the glue t! 50 ^ ! 100&quot; : 197±125 ,- I —— i 々: Due to differences in animal weight, it is about Dong. Example 30: Preparation of 5-CNAC | The preparation of 5-CNAC was similar to the conditions in Example 26. Example 31: freeze-dried sCT/5-CNAC sodium salt | 19.0072 g of 5-CNAC prepared in Example 30 and sCT cold prepared in Example 19; Dongganyu powder and 4853⁄4 liters of 0.2 N NaOH in steam Mix in the bath. The final volume is 505 ml. Preparations 187, 138, 74 and 160 ml of four batches contained 5-CNAC #3 salt and 28, 48, 40 and 360 mg of sct, respectively. The approximate amounts of 5_CNAC sodium salt are 7, 5, 2.5 and 4.5 grams, respectively. Example 32: Administration of sCT/5-CNAC sodium salt oral delivery of mice | Method 5 according to Example 21 and standard procedure for the EIA kit. The mice were orally administered a capsule containing 13 mg of Example 3 in the middle of four j ' ? ί

1282783 V. INSTRUCTIONS (49) | One of the batches of lyophilized powder, with 1 ml of water. The dose and administration results of sCT/5-CNAC i sodium salt are shown in Table 1. Table 10: Mouse sCT/5 -CNAC sodium salt oral delivery agent dosage form SCT/5 -CNAC sodium agent mann i sCT agent f ί (mg / kg) | (grams / kg) i ! ! Average crest! Serum sCTtSD ! (pick / soar) 1 〇 5a) capsule 丨 | 50* | · loo*· S 125 153 (15b) capsule: 50&quot;; 400^ 1 178±35^ (15c) capsule | 50&quot 1 800^ Γ 546±586 i(15d) plastic 曩! 50^ | 4000* | 757±1234 : Due to the difference in animal traits, it is about c; * I The above patents, publications, applications and test methods are uniform And included in the reference jj literature. Anyone who is familiar with this technique may make changes or changes with reference to the contents of the above description. Therefore, the present invention is based on the scope of the patent application, - ;

1071-3452-PF.ptd第57页

Claims (1)

1282783 Case No. 891207 Deng Liu, the scope of application for patents η &lt; Japanese PCT is a carrier of the two sodium salt, Li Chuan Er Er 1 The following chemical formula 公告 bulletin R2, where 'r, R2, R3 and R4 independently Is hydrogen, 0H, NR6R7, dentate, alkyl, or C plant c4 alkoxy, · 1 妒, is an unsubstituted C2-Cl6 alkylene group, unsubstituted c2_Ci6 alkenylene group, substituted [Cl- Cl 2 alkyl (arylene)] or unsubstituted [aryl (CrQ alkylene)]; and R6 and R7 are independently hydrogen, oxygen or Ci-q alkyl. 2. The disodium salt of the transfer agent of claim 1, wherein the transfer agent is N-(5-chlorosalicylate)-8-aminooctanoic acid. 3. The disodium salt of the transfer agent of claim 1, wherein the transfer agent is N-(1〇[2-hydroxybenzylidene]amino)decanoic acid. 4. The disodium salt of the delivery agent of claim 1, wherein the delivery agent is sodium N-(8-[hydroxyoxybenzylidene]amino)octanoate. 5. The ethanol solvate of the disodium salt of the transfer agent according to claim 1, wherein the transfer agent is N-(5). - chloroauric acid 8-aminooctanoic acid. Page 58 l〇71-3452-PF3.ptc 1282783 Emperor 891207 Deng Liu, Scope of Patent Application 曰 Amendment 7. The ethanol solvate described in the scope of claim 2, wherein the transfer agent is N - ( 1 〇 - [2. Hydrogen nitrogen 8] The ethanol solvate according to claim 5, wherein the transfer agent is sodium N-(8-[hydroxyoxymethionyl]amino)octanoate. The monohydrate of the di-salt salt according to the first aspect of the invention. The monohydrate according to claim 9, wherein the transfer agent is N~(5 - gasified salicylate ) ~ 8 - Aminooctanoic acid. II. The monohydrate according to claim 9, wherein the transfer agent is 1 〇-[2-hydroxy oxomethylamino) decanoic acid. The monohydrate of claim 9, wherein the delivery agent is sodium N-(8-[hydroxyphenyl]amino)octanoate. 1 3 · a composition comprising at least 5 〇 % of the disodium salt described in claim 1 of the patent application, based on the carrier and salt of the composition of 100 ° /. The composition according to claim 3, wherein the composition is at least 90% of the disodium salt, based on 100% of the total weight of the transfer agent and the salt in the composition. . 1 5 · a composition comprising: (a) The disodium salt of the transfer agent of claim 1, or the (b) at least one active agent. The composition of claim 15 which comprises at least 5 % by weight of the disodium salt based on the transfer agent and the salt of the composition 1 〇 〇 Z 1071-3452-PF3.ptc Page 59 1282783 _ Saizhong 89120728 _Year ^___§-Fresh-_ VI. Patent application scope Total weight. 1 7 · If the application for the 'Li Fan' circumference of the '1 technology, less than 90% of the disodium salt, based on the total weight of the transfer agent and salt in the composition of 1 〇〇〇 / 0. The composition of claim 5, which comprises at least 90% of the monohydrate based on the total weight of the hydrate of the carrier disodium salt in the composition of 100%. The composition of claim 15 wherein the active agent is selected from the group consisting of: growth hormone, human growth hormone, recombinant human growth hormone (rhGH), and cattle growth Mongolian, pig growth hormone, release of the growth of the hormone of the hormone 'interferon, including interferon, interferon, interferon, interleukin 1, interleukin 2, insulin, porcine insulin, bovine insulin, human Insulin, human recombinant insulin, insulin-like growth factor, IGF-1, hepatic cadherin, unseparated hepatic condensate, hepar 1 no ids, chondroitin, cartilage, low molecular weight hepatic calcin, Very low molecular weight hepatic calcin, ultra low molecular weight hepatic calcin, calcium regulating hormone, shoe calcium regulating hormone, salmon calcium regulating hormone, porcine calcium regulating human calcium regulating hormone, red protein, atrial naturetic", factor antigen, Monoclonal antibodies, body hormones, protease inhibitors, gonadotropin, gonadotropin-releasing hormones, oxytocin, = hormone release, HOOL, Mengyi, stimulating serotonin, cerebral glycosidase, coagulation Whitening, Whale's blood, prostaglandin, cyclosporine, angiotensin, linger, kegeling sodium or disodium salt, sinkamycin, desferrioxamine, parathyroid hormone, parathyroid hormone fragment , antibacterial agents, including antifungal agents 1071-3452-PF3.ptc Page 60 1282783 A similar substance of such a component, a tablet p, a mimetic or a modified derivative thereof, or a mixture thereof. 2. The composition of claim 5, wherein the active agent is selected from the group consisting of hepatic cadherin and calcium regulating hormone. 21. A pharmaceutical composition for promoting delivery of an active agent, comprising: (a) the composition of claim 5; and (b) (i) - a excipient; a diluent; (111) a disintegrating agent; "ν" - lubricant; (v) a plasticizer; (Vi) a colorant; (vii) - a carrier for administration; or 'V11i' or a mixture of any of the following: • a solid used to promote delivery of the active agent A formulation for administration comprising a cold-blown #'/east dry mixture comprising: the di-nano salt as described in claim 1; and (b) the active agent. 2 3 · A composition as claimed in claim 15 is used for the preparation of animal liver pacingin as described in claim 15 of the patent application. The method for preparing an anhydrous di-nanopharmaceutical of a transfer agent is to dry the transfer agent, an ethanol solvate of a sodium salt, wherein the chemical formula of the transfer agent is as follows: 1071-3452-PF3.ptc Page 61 1282783 Correction R1 κ. , ΟΗ Υ ο where R1, R2, R3 and R4 are independently 氲, -OH, -NR6R7, dents/, C: -C4 alkyl, or C4 alkoxy; unsubstituted H6 Affiliation, unsubstituted C2-Cl6 alkenylene, I [Ci - Ci2 alkyl (arylene)] or unsubstituted [aryl (CC)); and R6 and R7 are independently hydrogen, oxygen Or Ci - ^ alkyl. 2 5 · Preparation of the transfer agent disodium salt ethanol solvation comprises:, method, a) the transfer agent is dissolved in ethanol to form a transfer agent / ethanol soluble sub-liquid; to - (b) transfer agent / ethanol The solution is mixed with a molar excess of sodium-containing ethanol solvate, wherein the structural formula of the transfer agent is as follows: , ΟΗ </ RTI> <RTIgt; Or, Ci-, alkoxy; R5 is unsubstituted c2-c16 alkylene, unsubstituted c2-c16 alkenylene, unsubstituted [Crc12 alkyl (arylene)] or unsubstituted [ Aryl (Ci-C12 alkylene)]; and R6 and R7 are independently hydrogen, oxygen or G!, C4 refining. The following steps: step (c) ethanol formation from step (b) An ethanol solvate is obtained in a solution of a solvate. . . : . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Obtaining an ethanol solvate of the transfer agent di-nano salt; (b) drying the ethanol solvate of the transfer agent di-nano salt to obtain an anhydrous disodium salt thereof, and '," (c) obtaining the anhydrous disodium salt a hydrate thereof, wherein the structural formula of the transfer agent is as follows: N^RSr〇H rT Mozhong' R1, R2, R3 and R are independently hydrogen, -OH, Yili 6^7, _ I 1071-3452-PF3.ptc Page 63 1282783 Case No. 89120728 Sixth, the scope of application for patents, C-C4 alkyl, or C!-c4 alkoxy; R5 is unsubstituted C2-Ci6 alkylene, unsubstituted (^ c unsubstituted [Ci-C] 2 alkane (Alaryl)] or unsubstituted [Aryl (poly, alkyl)]; and 1 to k sub-R6 and R7 are independently hydrogen, oxygen or q-c4 alkyl. The second salt, the transfer agent has the following chemical R4 Ο 丨s /OH Y 〇 wherein R1, R2, R3 and R4 are independently hydrogen, -OH, -NR6R7, _, Ci - C4 alkyl, or C! ~ C4 alkoxy; R5 is unsubstituted or a C2_CH alkylene group, a G-Cu alkenylene group, a [Cl-Cl2 alkyl group (arylene)] or an aromatic group substituted with one or any combination of _, hydroxy, C-G-alkyl or q-C4 alkoxy The group (cQ2 alkylene group)]; and R6 and R7 are independently hydrogen, oxygen or C-wide. 2. The composition of claim 15, wherein the active agent is a calcium regulating hormone. 3〇.^tηψψι^„2g^ # mJ ^ tm# The agent is N-(5-chlorinated acid)_8_aminooctanoic acid. 31. The composition as described in claim 5 of the patent application In which activity 1071-3452-PF3.ptc Page 64 1282783 1071-3452-PF3.ptc第65页