SA02230168B1 - A pharmaceutical formulation comprising an iota-carrageenan compound and at least one neutral gelling polymer. - Google Patents

Abstract

Abstract: The present invention relates to an oral "pharmaceutical" composition comprising an iota-carrageenan compound, one or more neutral gelling polymers, and a pharmacologically active and active base component, which is the composition which inhibit the release of the pharmacologically active base ingredient from a formulation at an acidic pH. This invention also relates to a process for manufacturing the aforementioned formulation; And the use of such a combination in the manufacture of medicine, , 1 1 Fig

Description

Y — _ pharmaceutical formulation comprising iota-carrageenan and at least one neutral gelling polymer Full Description

background of the invention

The present invention relates to a new oral pharmaceutical formulation; It contains an active ingredient

A major pharmaceutical whose solubility depends on its pH; He who discourages

Release of the main pharmaceutical active ingredient from the formulation at an acidic pH (preferably 0 pH less than “) and preferably provides a highly dependent controlled release

The pH of the active ingredient is pharmaceutically controlled over a wide pH range in the channel

digestive a process for manufacturing said formula; And use the formula mentioned in the treatment.

Active controlled-release pharmaceutical formulations are pharmaceutical products

It is desirable as it allows Lay the following: to improve drug treatment; In addition to the possibility of reducing the times of taking 0 dose and reducing unwanted side effects. Only the design of such systems that are controlled

In launching it, it is not an easy matter; Especially when the goal is to make a pharmaceutical formula

For oral administration and must pass through the gastrointestinal tract; from what appears; among some

Other features include a wide variation in pH along its length.

Many drugs that exhibit basic properties ionize at low pH and become ALE 1 highly soluble in the said pH region compared to neutral environments.

the other. The phenomenon of solubility according to the pH in the gut; maybe

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Y — — lead to multiple forms of drug release; With the emergence of some parallel problems of vital activity in the body of the organism. Several attempts to overcome the pH-dependent solubility problem of basic drugs have been described. These methods include the use of an integrated polymer; which cannot be dissolved at such a low pH; To disrupt drug release in low pH periods (see eg: and A.

Streubel eral.

J.

Controlled Release, 67,101-110 (2000), 1154968508 “or a low molecular weight organic acid is incorporated to make a low acidic environmental pH within the formulation group; And then keep the solubility potential of the drug constant (al) for example: K.E.

Gabr., Eur.

J.

Pharm.

Biopharm., 38(6). 199-202 (1992), and V.K.

Thomas and The. 01 Zimmer, Pharm.

Ind. 51_(1), 98-101 (1989) Inclusion of an ionic polymer (eg sodium alginate) showing pH-dependent solubility in a drug formulation that also includes a neutral polymer; It gives the properties of an insoluble gel at a low pH; Which leads to a diffusive barrier that was discovered to be the main LN 10 that works to disrupt drug release at low pH: Pharmaceutical De-velopment and Technology, 2(1), 25-]am as adsorbent 1.1 and [: 1054792452 (1997) 31] and other methods that have been used that use the loaded polymer to affect the release of the drug, whether by ionic interaction with the drug, see:

— ¢ —_ 9

C. Caramella et al. Pharm. Res. 14(11), 531 (1997), H.Y. Park et al. Drug Delivery. 513-18 (1998), N. Caram-Lelham, Ph.D. thesis, Uppsala University (1996) or by affecting the gel properties and volume increase of these polymers (see Picker, Drug Dev. and Ind. Pharmacy, 25(3) 339-346 (1999) 121/4). The 0 used here generally depends on the type of polymer Bavejaetal (1987) 39, 45-39 reveals that when nonionic polymer (HPMC) is mixed with anionic polymer (NaCMC) the release is delayed. : Ranga Rao et al, Drug Dev Ind Pharmacy, 14, 2299 (1988) mixtures of methylcellulose and NaCMC result in different release forms, mixtures of lambda-carrageenan and active ingredient 0 are disclosed in the application International 99/716785. Method combinations for providing a non-pH-dependent release are mentioned (see IS Nos. 757/17 // AT, 44/143065 and 184/99). Polymers with different water solubility and different swelling properties; the formulation of each can vary 0 + adjust these properties to provide tunable release rates of 0. Clear pH-dependent solubility such as sodium alginate ¢ and a gelatinous polymer with a low pH-dependent solubility Jia hydroxypropyl methylcellulose (HPMC) or polyethylene oxide. Uses either fully encapsulated polymer; As the copolymer of methacrylic acid in International Application No. 779717/56 or Amla:

_m — Lf EUDRAGIT® or 5 which form certain types of methacrylic acid polymers in . IS 749705 / 94 or water soluble polymer; Fie ethyl cellulose in International Application No. 4784/94. However, these Gok were generally not essential target drugs; It also relied on the type of all-coating or cell-insoluble polymer such as 0-methacrylic acid polymer; or pH-dependent gel polymer; such as sodium alginate; To disrupt drug release in low pH environments at least partially. Description: BONFERONIM.C.

ET AL. in 'On the employment of lambda-carrageenan in matrix system.' II. ] ambda-Carra-geenan and hydroxypropylmethylcellulose mixtures' JOURNAL OF CONTROLLED RELEASE vol. 30, 1994, pages 175 - 182 \ A formulation containing HPMC j lambda-carrageenan, but it failed to mention Iota-carrageenan. US Patent No. 797,457 does not mention any of them; A- Or European Patent No. Ty — cordon Or Japanese Patent No. 87149719-A Or International Application No. 971885-a Iota-Carrageenan. Vob reveals: HAM-YONG PARK ET AL in “Effect of pH on drug release from polysaccharide tablets’ DRUG DELIVERY vol. 5,1998, pages 13-18 da

for formulations of tablets containing sodium alginate or Iota-carrageenan; However, he did not succeed in

He mentioned the use of neutralizing gel polymers.

General description of the invention

The invention provides all 385 oral pharmaceutical formulation comprising Lad-carrageenan

0 and one or more neutralizing gel polymers and a basic pharmaceutical active ingredient Cua that denatures these

The formula releases a key latent active ingredient from the formula at an acidic pH

(Preferably less than oF pH, especially at pH 1.)

Essentially a pH release means that the release rate is greatly disrupted at the number

pH 1 and increases or is unaffected at pH VA hence the amount of Pharmaceutical 0 Principal Active Ingredient released at any time becomes less pH dependent.

The present invention additionally covers an oral pharmaceutical formulation comprising

Iota-carrageenan, one or more neutralizing gel polymers, and a pharmaceutically active ingredient.

Preferably, Iota-carrageenan is present in the formulation of the invention at a level of more than

- Of natural origin. One of the grades of Iota Slat lS - by weight. Preferably IOTA 70 having a viscosity (FMC can be obtained from Biopolymer A) Pharmaceutical Carrageenan 1

Do not exceed ©cp and preferably range from © = 01cp (from *,07 of the solution

which is heated to 87°C; The viscosity at VO °C is then measured using

The Brookfield LV viscometer with #1 spindle operates at SAY speed

minute). Type of technical degree for IOTA - Carrageenan (which can be obtained from

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- a - Fluka Biochemica (preferably a viscosity of not less than VE MPa per second for a Sled solution heated to 10°C; after which the viscosity was measured using a drop-ball viscometer; type (Haake used with C3 Lauda and Hakke Mess-System III thermostats and using stainless steel balls coated with gold with a density of VA 0 gsm). Natural gel polymer is one or more polymers of mixtures of one or more corrosive polymer(s) having gel properties and with solubility mainly dependent on pH.Natural gel polymer is preferably present in the composition at a level of not more than 107 and preferably not more than 7#1 by weight (indicates ALE corrosion” and “corrosion” lead to dissolution or decomposition, either alone or in combination. Solubility can be improved by mixing; decomposition can be improved by mechanical interaction with a solid). Suitable gel polymers include polyethylene oxide (PEO) derivatives. ) and members of the PEO family (such as polyethylene glycol (010); preferably naturally occurring in solid Ala; with appropriate molecular weight or viscosity). So the natural gel polymer for example is polyethylene oxide 1 . or polyethylene glycol. In the case of using a single neutral gel polymer, it is preferable that the molecular weight of PEO be greater than ; m(;molar) (ie, the molecular weight is greater than − to +m); Corresponding to an aqueous solution viscosity in the range of 1650 - 0000 MPa (or V0, = 000 cc measured in an aqueous solution of #1 at Yo °C; using a 0 -_ Brookfield RVF two-column viscometer speed?rpm). Examples include Ed/A

M -

Others for PEO are those in which the molecular weight is approximately © million (0 mol); corresponds to range

viscosity of an aqueous solution in the range of 55.00 - 1500 MPa; Or a PEO molecular weight of

Approximately A million (A) molar) corresponds to an aqueous solution viscosity range of 105.00 = 000080 M

Pascals per second. This weight covers the typical viscosity of the solution (in centi-Poise) measured at m YO °C. And used for this polymer in:

USP 24/NF 19, 2000 edition, pp. 2285-2286. Hence PEO can have weight

Molecules ranging from 4 A= million.

If PEG uses a single neutral gel polymer it is preferable to have a molecular weight of “aS” as

Its molecular weight is approximately Yeon and corresponds to a viscosity range of Youu - 77000 0 MPa (or 77060 = Your centipeas) measured using an aqueous solution (coger/wt)

at 01°C; using an Ubbelohde capillary viscometer (or equivalent) [ Ref:

,. [European Pharmacopoeia 3rd Ed., 2000, Supplement, p. 908-909

Other suitable gel polymers include cellulose derivatives such as hydroxypropyl methylcellulose

Cellulose (HPMC) or Hydroxyethylcellulose (HEC) (but preferably HPMC with 1 of suitably high viscosity (eg 110140 00001cP) or HPMC

000 cents ls type HH HEC or type HEC 11). when used as a natural polymer

single Hydroxypropyl Methyl Cellulose Copolymers 00001 HPMC ie centipars and

- 75050 centipores having respectively apparent wives of the amount of 105001 HPMC

5 MPa (or Vous - 1406060 cP) and 70001 - 117860 MPa 03 (or 70001 - 11701 cP) when measured at Yo 77 °C in an aqueous solution of 77 (w/

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f - weight); and its calculation by reference to the dry matter; using an Ubbelohde (or equivalent) capillary viscometer 0 of a type of hydroxyethylcellulose polymer; Example 250 Natrosol “Pharma Type HH” from Cua Hercules Incorporated (Aqualon) shows a typical Brookfield viscosity of approximately 560,000 MPa using the Brookfield Synchro-Electric Model LVF tool. oe by Gel conditions) the concentration of the solution; by column number of and by a spindle speed of 0” rpm; and a Yo factor of 7000 °C (see:

Natrosol Physical and Chemical Properties booklet, 33.007-E6 (1993), p. 21 When a mixture of natural gel polymers is used this mixture may include for example a mixture of two or more PEO or two or more HPMC or HPMC 5 PEO or PEGS PEO 0. Example PEO is a molecular weight; or © or A million can be mixed with PEO with a molecular weight of 1 million; PEO with a molecular weight of PEO chee, ove by weight 000001 Aa or

0 molecular weight Seve

instead; The natural gel polymer (example 0:31) can be used in combination with

A natural, non-gelatinous polymer (eg low molecular weight PEG Jie 226 has a molecular weight less than 000011 1). An example of low molecular weight PEG in such combinations is PEG which has

Molecular Weight 8000 (corresponding to a viscosity range of 760 - 001 MPa) or PEG wt.

Molecular 0001 (corresponds to a viscosity range of 7700 = 7700 = 000 MPa).

A mixture of HPMC grades can include both low viscosity (non-gel)

And high viscosity (gel). For example 50 110140 Jal cp V0 hpmey 0 cp and ©110140 1 cp) have an apparent viscosity of 01-460 MPa;

- Ve = 70.1 - 1" MPas; 8, - 7.7 MPas; according to the method previously specified may be used as mixtures with 00001 centipoise HPMC or HPMC . Sainte Boise 1 © . In a mixture; the higher the molecular weight of the PEO used the less of that polymer is needed to make the composition according to the invention. The specific composition of the present invention depends on the molecular weight and its distribution. 0 In one particular aspect Hereby the neutral gel polymer is PEO with a molecular weight of approximately aly; million or more or PEO with a molecular weight of approximately 700055 or greater or a cellulosic derivative having an apparent viscosity of approximately 7950 cP or greater (measured as Percentage of neutralized gel polymer (eg PEO or PEG or HPMC and especially PEO or HPMC or mixtures thereof with each other or more of PEO or HPMC (for Iota-carrageenan) 01 to 400 : 0 (particularly from A : 10 to 01 : 80 preferably in the range from vo

Ao 0 : 50 eg approximately fe to the basic pharmaceutically active ingredients having one or more principal groups which 1 to 1 (especially 01 to 1 (eg from VY to 1 preferably in the range of its pKa being greater than its pKa and optionally also having one or more of its principal groups)

11 - - from .01 and then; The main pharmaceutically active ingredient can have one or more pKa values but at least one of them is from 1 to VY (eg from 1 to 01 (and especially from 1 to 7). Examples of base groups in these pharmaceutically active basic ingredients having a pKa ranging from 1 to 11 (Je) for example 1 to (Ve) include 0-hydroxyamidines; secondary and tertiary amines or primary and secondary amides.

Suitable basic pharmaceutical active ingredients should preferably have low to moderate water solubility (eg water solubility maximum 00 mg/mL ali) 00 to 10 mg (Jaf at YO °C die (pH 7.0); They are positively loaded with one or more positive charges (depending on the pKa number of 0 base groups present in the pharmaceutically active ingredient) to a low pH le) way

The example is a pH of 1 to 7 (especially a pH of 1 to ). An appropriate pharmaceutically active base ingredient is for example an ingredient with cardiovascular activity (eg a peptide, a cama-like or a thrombin inhibitor). Peptide thrombin inhibitors have a molecular weight of less than 0001 and have 1, 7, © or ; of the peptide and it exhibits a solubility potential of 1 that is pH-dependent. They include the peptide thrombin inhibitors (and prodrugs thereof) mentioned more generally and more specifically in Claesson in Blood Coagul. Fibrin. 5, 411, (1994) and further disclosed in US Patent No. 8 and US Patent Applications WO 97/23499, WO 97/02284, W097/46577, WO 9S/01422 , WO 93/05069, WO093/1115 WO 95/23609, W095/35309, WO WO 94/29336, WO 93/18060 WO 95/01168 0, 96/25426, EP No. 623,596

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364 002 362 212 195 525 542 877 526 390 185 779 641 046 559 231 468 780 648 459 601 317 669 642 686 881 293 167 530 344; and peptide thrombin inhibitors (or prodrugs thereof), especially those comprising HOOC-CH2-} inogatran, melagatran YI - YIN (pula—tRCgl-Aze-Pab-H [[[[ ; (aminoaminomethyl) ) phenyl]-methyl[amino] 0 carbonyl]-1-azetidinyl]-1-cyclohexyl-¥-oxoethyl] [YD 7118 8]- ) f(zymegatre n<EtO2C -CH2-RCgl-Aze-Pab-OH See example VY from patent “WO 97/23499 glycine;17-1[-cyclohexyl-¥-[?-[[[[;-[( hydroxyamino)aminomethyl] - phenyl] - methyl [amino] carbonyl] - 1-azetidinyl] - ?-oxoethyl] - ethyl ester [8 - R¥)*8)]- ). 0 On the other hand, peptide thrombin inhibitors (or prodrugs thereof) include inogatran; Malagatran » 5 H376/95 Ph(3-CI)(5-OCH,CH,F)-(fl)CH(OH)C(O)-Aze-Pab(OMe)and Ph(3-CI) (S-OCHF,)- (f)CH(OH)C(O)-Aze-Pab(OMe). In another aspect of the present invention a formula as given herein is provided in which the active ingredients 1 The basic pharmacology is Ph(3-CI)(5-OCHF,)-(fl)CH(OH)C(0)-(S)Aze- Pab(OMe) {Compound A}; Ph(3 -CI)(5-OCHF,)-(fl)CH(OH)C(0)-(S)Aze-Pab(2,6- diF)(OMe) {Compound D}; \YoY

Ph(3-CI)(5-OCH,CH;F)-(fi)CH(OH)C(O)-(S)Aze-

Bab(OMe) {Compound E}; Ph(3-CI)(5-OCHF,)- (f)CH(OH)C(O)-(S)Aze-Pab(OH) {Compound F};

Ph(3-CI)(5-OCHF,)-(fl)CH(OH)C(O)-(S)Aze-Pab(2,6- diF)(OH) {Compound G}; Ph(3-CI)(5-OCH,CH,F)- ° (fi)CH(OH)C(0)-(S)Aze-Pab(OH) {Compound H}.

FandH The compound © may be prepared by methods similar to those mentioned below for the preparation of compounds In another aspect of the present invention a pharmaceutical composition is provided in which the basic pharmaceutically active ingredient is: 1,4- ( {3- ]7- (3, 3-) dimethyl-2-oxobutyl)-9-o0xa-3, 7-diazabicyclo [3,3.1] non-3 yl] 01 propyl } amino)benzonitrile (B (hereinafter referred to as compound 2.tert-butyl2- {7-[3-(4-cyanoanilino)propyl]-9-oxa-3, 7-diazabicyclo-[3.3.1]non-3-yl} ethylcarbamate 3.tert-butyl 2-{7-[4 - (4-cyanophenyl) butyl]-9-oxa-3, 7-diazabicyclo- [3.3.1] non-3 yl} \o ethylcarbamate or

_— $ \ _— tert-butyl 2- {7-[ (2S)-3- (4-cyanophenoxy)-2-hydroxypropyl]-9-oxa-3, 7 diazabicyclo .4 non-3-yl } ethylcarbamate [3.3.1] (where this compound is referred to hereafter as compound (C). These compounds were then explained in the international application co / YA44Y 0. On the AT side, the basic pharmaceutically active compound is metoprolol or its aie salt (such as succinate or a tartar thereof).The formulation of the present invention may include a curing additive; a stabilizer or plasticizer; a coloring matter; a lubricant (Jia (sodium stearyl fumarate) and a binder or filler or a surfactant or other excipient for use in pharmaceutical preparations. 0 In one aspect of the formulation of the present invention includes a lubricant (such as sodium stearyl fumarate) 0 In another aspect of the present invention the molar ratio of Iota-carrageenan of a pharmaceutically active ingredient ranges from ? : 1 to .0 : 1 Another aspect of the pharmaceutical formulation of the present invention comprises 11-77860 of Iota-1 carrageenan. The AT side of the pharmaceutical formulation of the present invention comprises 15 - 7850 of one or more neutral gel copolymers. the

- 1 and

In another aspect the pharmaceutical formulation of the present invention comprises of the active odie-1

Basic pharmacist.

In another aspect, the pharmaceutical formula of the present invention includes from zero -

0 (especially from eV - 1 the active substance of the treatment; or the substance of the fixation;

0, plasticizer, colorant, lubricant, binder, filler or other excipient

It is used in pharmaceuticals.

The mechanism behind inhibition of the release of the pharmaceutically active ingredient from the formulation of the number

acidic pH (particularly pH-dependent release) which is

which is believed to be as follows. at low pH; A pharmacologically active basic ingredient 0 is expected to have a high solubility as it is in a strong ionic state;

Therefore call will be expected to be fired faster than any set of neutrals. from

Theoretically, Laie is acidic pH and in the presence of Iota-carrageenan; And there is

ionic attraction between the positively charged iota-carrageenan and the positively charged drug; And who

It hinders the release of the drug and thus contributes to a more stable release. when the pH is higher; ve i.e. Laie the pharmaceutical drug is less ionized or not ionized at all; Hence it is expected

It appears on slow release from any neutral group; It is theoretically an ionic reaction

The aforementioned is less important, and the form of release can be largely controlled through

associated swelling, gelation and corrosion of neutral gel polymer(s) and anionic polymer; And

The Iota-Carrageenan used in the formula.

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ultimate swelling properties; The gelation and corrosion of the formulation presented in the present invention are dependent

on some properties such as molecular weight; The molecular weight distribution of a polymer or polymers

jellies and anionic polymers. This is also associated with pH-dependent decomposition

of the anionic polymer. Hence the different release rates v by the active ingredient

0 Pharmacologically it can be obtained by denaturing the gel polymer Je) eg weight

molecular weight distribution) of the gel polymer; How much Iota-carrageenan is in

The formula and/or ratio of the gel polymer to Iota-carrageenan.

The composition of the present invention may exist as a solid dosage form (such as tablets, capsules, or

pellets or powder to be spread into a suitable container; or as a multiple formulation (eg, coated pellets given in ¢ tablets, capsules, or scented powders).

In one aspect of this invention a tablet comprising of Orr = Yo mg is provided

(particularly from 1 - 401 mg of a basic pharmaceutical active ingredient (Jia) 11376/95 or compound EE or

(C R 8

When the pharmaceutical formulation of the present invention is contained in a tablet it is preferable to vo the tablet so that all of the base pharmaceutical active substance is released; in ionic or non-ionic form

Based on the pH of each part of the gut over a period of time of approx

0 hours for example from YY - 18 hours (or alternatively from 01 to YO hours)

In another additional aspect, a special process is provided for preparing a formulation of the present invention that includes:

Mixing Iota-Carrageenan; and one or more neutralizing gel polymers and a pharmaceutically active ingredient

_ 7 \ main ham; Optionally incorporating said mixture (preferably in the presence of a lubricant (such as sodium stearyl fumarate, sold under the trade name M) to form the tablet. The tablet formulation may be prepared eg by direct pressing or wet-pellet techniques. - Usd for Technologies Direct pressing A basic pharmaceutical active ingredient is mixed with gel polymer, 0-carrageenan and additional excipients as required Lubricant (such as sodium stearyl fumarate) is sifted and added to the Iota-carrageenan mixture and further mixing is carried out. It compresses the resulting mixture into tablets. For wet pellet technologies, the basic pharmaceutically active ingredient is mixed with a gel polymer and

Jia) Iota-carrageenan. The resulting mixture may then be wetted with: a solution of suitable binder 0 dissolved in a suitable solvent (such as ethanol or water); or a suitable polyvinylpyrrolidone (PVP) solvent (eg ethanol or water) and the resulting mixture is granulated using standard or modified granulation procedures (eg spray granulation). After drying the resulting granules (for example in an hour) YE - 01 Jia) oven and at a suitable temperature (eg .5 degrees Celsius) for a suitable period

Jia) grinding of granules (eg dry or wet grinding); mixing it with a lubricant, sodium stearyl fumarate; magnesium stearate or talc) and the resulting composition is pressed into tablets. Dry granules can be used to fill capsules (such as capsules made of gelatin). In another aspect, the present invention provides a process for preparing a formula as mentioned before. yoy

1 - The active compounds of thrombin and their prodrugs can be used for the treatment and/or prevention of hypercoagulable thrombosis in the blood and/or tissues of animals including humans. It is well known that hypercoagulability may lead to occlusive disease. The conditions associated with hypercoagulability and vascular occlusion which can be mentioned can be present or obtained© from protein resistance©; such as factor V mutations (factor Leiden V), hereditary or acquired defects in antithrombin IT, protein ©, protein 8, and heparin cofactor [1]. Other conditions known to be associated with hypercoagulability and vaso-occlusive disease involve circulating antiphospholipid antibodies ( anticoagulants in lupus); homocystinemia heparin inducing thrombocytopenia and fibrinolytic defects as well as thrombotic symptoms (eg diffuse cardiothrombosis (DIC) and general vascular injury (eg those resulting from surgery) jig Another aspect of the invention is Mall formulation as mentioned above for use in treatment (whether preventive or curative) eg as a treatment (eg treatment of cardiovascular disorders; Jie thromboembolism). According to the present invention it is useful for the manufacture of a drug for use in the treatment.Another aspect of the present invention provides for the use of the present formulation for the treatment of a cardiovascular disorder (eg vascular occlusion) in warm-blooded animals suffering from or at risk of this disorder.Some inhibitors Thrombin peptides or terminated prodrugs can be prepared through the methods described below. \Voy

- 11 -

General procedures

TLC was performed on silica gel. JS HPLC analysis was performed using £7 mm You X

mm of Chiralcel OD with © cm of shaft guard. The column temperature was maintained

at YO °C; and a flow rate of 10 mL/min that was used. A Gilson UV 115m detector at YYA nm was used. be the mobile phase of

hexanes; ethanol, trilauroacetic acid, and the appropriate proportions are listed for each compound.

likewise; The product was dissolved in the minimum possible amount of eschaol and this resulted in phase dilution

mobile.

In the preparations mentioned before; LC-MS/MS was performed using an HP-1100 instrument equipped with a 0 “Tm© 3 CTC-PAL” injector; 001 x mm for ThermoQuest, Hypersil BDS-C18 Column and Explorer

API-3000 (Sciex) MS has been used. The flow rate was 0.7 mL/min as phase

moving (gradually) from 01 - 7490 acetonitrile with 0 = 10m; mM of acetate

aqueous ammonium; Each contained 0.7 Gal formic acid. Otherwise, a spectrum is registered

Low Micromass (LRMS) using Micromass ZQ mass spectrometer at ESI posneg 1 ion phase shift (mass range 001 - 8080); and high resolution mass spectrometry (HRMS).

Recorded with a Micromass LCT spectrometer in 5 negative ionization phase (mass range m/z

)0001 — 0 with Leucine Enkephalin (CosHaNsO7) as an internal mass standard.

The HNMR spectrum was recorded using tetramethylsilane as an internal standard.

mail

YY. = - Preparation of Ph (3-Cl) (5-OCHF)-(R)CH (OH) C(0)- (S) Aze-Pab (OMe) (compound (A ,0 Nf) N-OCH, "9 HO 5 N NH, 8 : Ci OCHF, (i) ¥ -chloro -© -methoxybenzaldehyde oF 0 -dichloroanisole added 1 /(4.0 g” £19 mmol) in THF (Yoo ml) 8 drip to magnesium metal (VEY) 0 g 0 mmol 0 before washing with 0” titer of Je 001) THF (HCI at Yo °C. After adding; 1 0-dibromoethane (3.5 g 1 !mmol) dropwise. Then heating the resulting dark brown mixture to reflux for day * hours The mixture was cooled to 0°C NON - diethylformamide (da 0) was added in one part The mixture was fractionated with diethyl ether (Je 4060 XY) + 0N was added of 5001 HCl ml).The organic extracts were washed with saline (yor ml) and dried (+250 H21); filtered and concentrated in vacuo to provide oil. Flash chromatography eluting (twice) on silica gel was performed using Hex: EtOAc ( ; ), 7.38 (s, 1H), 7.15 (s, 1H), (s, 3H). Vo 3.87 \VoY

1 - - (Nn) -chloro-0-hydroxybenzaldehyde solution of ©-chloro-©-methoxybenzaldehyde (YY,A) g; VTE mmol See step (i) above) in (Je YO) CHIC cooled to 0°C. Boron tribromide 1117 ca (15.8 mL) was added dropwise over 10 minutes. After stirring, reaction mixture 0 was stirred for 2 hours; water (00 Ja) was added slowly. The solution was then extracted b EO (#0 ml). The organic layers were collected; dried over sodium sulfate; filtered and concentrated in a vacuum. Flash chromatography eluting performed on silica gel using HextEtOAc (; : 1) To provide the compound given in the subheading (5,7 Yo al a 7). , 7.10 (s,1H), 3.68 00 (s,1H) (iii) ¥ -chloro-0-difluoromethoxybenzaldehyde solution of ?-chloro-0-hydroxybenzaldehyde (V,0) g A mmol see previous step (i) in 1 - propatol (YOu) mL) 5 (Je Vo) 160117700 heated to 1 _ reflux. while flipping; Bubbles of :011017 were made in the reaction mixture for two hours. The reaction mixture was cooled and acidified with 1 N of HCl and extracted with EtOAc (XY) 0 mL). The organic layers were washed with brine (Ja 001) and dried over magnesium sulfate), filtered, and extracted in a vacuum flash chromatogram by eluting using (1 : 4) Hex:EtOAc to provide the compound given in the subheading (6; ; g; x. 74:11 after

_ Y Y _ 'H NMR (300 MHz, CDCIs) 8 9.95 (s, 1H),7.72(s, 1H),7.52(s, 1H), 7.40 (s, 1H), 6.60 (t, 1. = 71.1 Hz , 1H)

Ph(3-CI)(5-OCHF+)-(ff,S)CH(OTMS)CN(iv) a solution of 1-chloro-0-difluoromethoxybenzaldehyde ) £1g; Je YY,Y mol ' 0 see previous step (iif) in :11:01 Yoo mL) cooled to 0°C. 1 A) Znl, grams added; 8.1 mmol) and trimethylsilyl A) Ala g; YV,4 mmol) was added and the reaction mixture was allowed to warm to room temperature with stirring for 0 h. the mixture was partially concentrated in a space of space to produce the compound given in the subheading as a liquid; which was then used directly in the next step (v) without further purification or 01 diagnosis. Ph(3-CI)(5-OCHFp)-(f1,S)CH(OH)C(NH)OEt (v) cal sa T,AY) Ph(3-CI)(5-OCHF,)- (f1,S)CH(OTMS)CN Assuming “,77 mmol; See previous step (iv) Added dropwise to 1101/2011 (+04ml). The reaction mixture was stirred oe al 1 hour; It was then partially concentrated in a vacuum to produce the compound given in heading Vo 1 as a liquid; They were then used in step (Vi) without further purification or characterization. Ph(3-CI)(5-OCHFp)-(fi,S)CH(OH)C(O)OEt (vi) Ph(3-CI)(5-OCHF,)-(fi,S)CH (OH)C(NH)OEt (4 1.7 g; assuming 77.7 mL see step (v) above) was dissolved in You. (THF ml). 0 M of (Ja £4 +) 1280+ was added and the mixture was stirred at 0 °C for 10 h; cooled and then concentrated 'YoY

yy - - partially in vacuum to remove most of the THF the reaction mixture was then extracted with (Ja 001 XT) ERO, dried with sodium sulfate and concentrated in vacuum to produce the compound in the subtitle as a solid; They were then used in step (vii) without further purification or characterization. (vii) 0 1,5(0-11)011(2)0(011)-(:11:)3-01()5-001117

z a solution of cal ja 1,Y0) Ph(3-CI)(5-OCHF,)-(fl,S)CH(OH)C(O)OEt assuming 77.7 mL Use see step (vi) previous in ? -Propanol (175 ml) and 17 (Safe You (KOH) ml) was stirred at room temperature for 11 hours. The reaction was then partially concentrated in space to remove most of the 7-propanol. The remaining mixture was acidified by 1 molar of 0,11:50, extracted with (Je 001 X ¥) ERO, dried 1182507, and concentrated in a vacuum to provide a solid. Flash chromatography on silica gel and eluting with MeOH : CHC, : 101,011 concentrate ( : * : 10 ) yielded an ammonium salt of the compound given in the subtitle. The ammonium salt was then dissolved in a mixture of EtOAC (VO) ml) and water (Je VO) and acidified using 1 N of 1101. The organic layer was separated, washed with vo-saline (00 (Je) and dried 1187507 and concentrate it in a space of space to provide the compound contained in

(vid) to (iv) steps 251 al ja ¥,Y) subtitle 'H NMR (300 MHz, CD;0D) 5 7.38 (s, 1H), 7.22 (s, 1H) ,7.15(s, 1H), 6.89 (t, Jy-F = 71.1 Hz, 1H). 5.16 (s, 1H)

\Voy

— p Y _ (viii) : Ph(3-CI)(5-OCHFp)-(ff)CH(OH)C(0)OH (a) and Ph(3-CI)(5- OCHFp)- (S)CH(OAc)C(O)OH (b) A mixture of ¥,Y) Ph(3-CI)(5-OCHF,)-(fl,S)CH) was heated OH)C(O)OH 11.1 g 1 mL 0 mol see step (vii above) 7.0-(lipase PS"Amano"s) in VY0 vinyl acetate) mL) (Je YYO) MTBE to EA reflux for 1 hour. The reaction mixture was cooled; filtered using celite and washing the filter paste using EtOAc, the filter material was concentrated in a vacuum by flash chromatography on sequential Ada all silica gel using NH4OH: MeOH: CHCI; concentrate (1 7 10:1) to produce salts Special ammonium 0 with the compounds mentioned in the subheadings (e) and (i).The compound (a) as a salt was dissolved in water, made acidic using 1 titer of HCL and extracted with EtOAc, the organic layer was washed with brine; dried with sodium sulfate; filtered and concentrated in space to provide the compound in subheading 1,Y) (a) grams; 1 7). For the compound given in subtitle 0 'HNMR(300MHz, CD;0D) 5 7.38 (s, 1H), 7.22 (s, 1H),7.15(s, 1H), 6.89 (t, Vo Hz, 1H), 5.17(s, 1H) 71.1 2g Ju mL

Yo - - Ph(3-CI)-OCHFp)-(CH(OH)C(Q)-Aze-Pab(Teoc) (iv) into a solution of (11( Ph(3-CI)( 5-OCHF)-(fi)CH(OH)C(O)OH 0.4 mmol see step (viii) above) H-Aze-Pab(Teoc) s (see international patent application 47064 eo] 0 g; VF) grams; 0176 mmol). The reaction mixture was stirred at 0 °C for 2 hours; Then at room temperature for another 11 hours. The reaction mixture was concentrated in a vacuum; Flash chromatography eluting was performed on silica gel (¥ times) first with (1 : 4) EtOH : CHCl then with EtOH : EtOAc (1 : Yr) and finally with CHCl : 11:011© (90 : 1) to provide compound contained 0 0 in subheading FV pl ja V0 (4) as a white solid. 'H NMR (300 MHz, 00:0, mixture of rotamers) 5 7.79-7.85 (d, J= 8.7 Hz, .Hz, 1H), 5.12 and5.20 (s, 71.1 gal 2H), 7.15-7.48 (m, SH), 6.89 and 6.91 (t, 1H), 4.75-4.85 (m, 1H), 3.97-4.55 (m, 6H), 2.10-2.75 (m, 2H), 1.05-1.15 (m, 2H), (s, 9H) MS (m/z) 611 (M+1)+ 0.09 M Ph(3-CI)(5-OCHFp)-(fll)CH(OH)C(0)-Aze-Pab(OMe, Teoc) (X) Ph(3 -CI)(5-OCHF,)-(fl)CH(OH)C(0)-Aze-Pab(Teoc) )+ ,+ g; 16 ml of filler (see step (ix) above); It was dissolved in 71 mL of acetonitrile and 50 g Toe (mmol) of ©0-methylhydroxylamine hydrochloride. The reaction mixture was heated at Ve °C for 2 hours. The solvent was also evaporated and the substance was partitioned between water and ethyl acetate. © The aqueous phase was extracted twice with ethyl acetate and the combined organic phase was \Voy Ju

_ Y 4 — 0.41 with water and brine, drying over sodium sulfate, filtering and evaporating. was the output. (#9 1) grams' H-NMR (400 MHz; CDCL): 8 7.83 (bt, 1 H), 7.57 (bs, 1 H), 7.47 μH) 2H), 7.30 (d, 2H) ), 7.20 (m, 1H), 7.14 (m, 1H), 7.01 (m, 1H), 6.53 (t, 1H), 4.89 (s, 1H), 4.87 (m, 1H), 4.47 (m, 2H ), 4.4-4.2 (b, 1H), 4.17-4.1 (m, 3H), 3.95 (s, 3H), 3.67 (m, 1H), 2.68 (m, 1H), 0 2.42 (m,1H) 0.97 ( m, 2H), 0.01 (s, 9H).

Ph(3-CIL)(5-OCHF.)-(fl)CH(OH)C(0)-(S)Aze-Pab(OMe) (xi) ml 1.17 g” +, € +) Ph(3-CI)(5-OCHF,)-(fl)CH(OH)C(0)-Aze-Pab(OMe, Teoc) allowed to react for TFA preceding); dissolved in ml of (x) mol; See step (water). Done NaHCO; 5 The residue was fractionated between ethyl acetate (TFA) a minute. Fey is vaporized. The aqueous phase was extracted twice with ethyl acetate and the combined organic phase was washed with water and brine and dried (sodium sulfate); filtering and evaporating it. The product was dried by collecting YA g. The yield was AEE necessary to make dala from water / acetonitrile. There were no (7 Ao). H-NMR (600 MHz; CDCI): 8 7.89 (bt, 1H), 7.57 (d, 2H), 7.28 (d, 2H), 7.18 (m, 1H), 0m ‎ 7.13 (m,1H), 6.99 (m, 1H), 6.51 (t, 1H), 4.88 (s, 1H), 4.87 (m, 1H), 4.80 (bs, 2H), 4.48 (dd, 1H), 4.43 (dd, 1H), 4.10 (m, 1H), 3.89 (s, 3H), 3.68 (m, 1H), da

2.68 (m, 1H), 2.40 (m, 1H). Compound preparation

D (Ph(3-CI)(5-OCHF3;)-(R)CH(OH)C(0)-(S)Aze-Pab(2,6-diF)(OMe)) D ° fluoro — ; — [(methylsulfinyl)(methylthio)methyl]benzonitrile (gla — 017(i) mL of 0 mol) dissolved in 0.04 (methylsulfinyl)(methylthio)methane) 7 mol degrees Celsius. Butyl lithium VA - dry in argon with cooling was added to THF

Yo mol) drip with stirring. The mixture was stirred for ½ YoU (1,1 Wee (Ja VT ) in 4-hexane; 0-trifluorobenzonitrile (0, μg; Foe) min. Meanwhile, the 0 °C solution was cooled. Celsius in argon VA = to Gla THF ml of 100 mmol) in oye YO

Yo and the previous solution was added through a medical tube to the last solution over a period of time of one minute; The cooling bath was removed and when the reaction reached room temperature it was done for 1 min. After removing the aqueous layer and extracting the remaining aqueous layer, THF is poured into 40,860 ml of water. The opaque ether layer was evaporated with water and dried on Jue sulfate three times with diethyl ether. Done 1. (#79:) gm Y sodium and its evaporation. B1] NMR (500 MHz, CDCI3) was 5 7.4-7.25 (m, 2H), 5.01 (s, 1H, diasteromer), 4.91 (s, 1H, diasteromer), 2.88 (s, 3H, diasteromer), 2.52 (s, 3H, diasteromer), 2.49 (s, 3H, diasteromer), 2.34 (s, 3H, diasteromer), 1.72 (broad, 1H) 'YoY

YA -— — gla — 1 1 (ii) fluoro - ; - formylbenzonitrile oY > = difluoro- ? [(methylsulfinyl)(methylthio)methyl]benzonitrile YOY) g; 78.37 mmol; see step (i) above) dissolved in 900 mL of THF and 3.5 mL of concentrated sulfuric acid was added. The mixture was left at room temperature for ? days and then it was decanted in £00 ml of water. This was then followed by three extractions using EtOAC, and the combined etherial phase was washed twice with aqueous sodium bicarbonate and brine, dried with sodium sulfate, and evaporated. Yield 1 1 V g A) 9 0 . Then work the position of the ida lg PC NMR photovoltaic group that showed the signal from the fluorine carbonate at 1167.7 ppm to show the proposed coupling model using two coupling constants in the order 770 Hz LY Hz on 0 arrangement in a manner that corresponds to the coupling Epso and meta of fluorine atoms. 1H NMR (400 MHz, CDCI3) 8 10.35 (s, 1H), 7.33 (m, 2H) gla — 1 oY (ii) fluoro- ; - 1-Hydroxymethylbenzonitrile - Difluoro — 4 — Formylbenzonitrile 1 ARY g; ANY ml 'Jee' see step (ii) above) dissolved in YO ml methanol; With cooling in an ice bath. 1 sodium borohydride (No 0 Ye.g » Ja A 1 01 mol) was added in parts with stirring and the reaction was left for 10 minutes. The solvent was evaporated and the residue was fractionated between diethyl ether and aqueous sodium bicarbonate. The etherial layer was washed with more dihydrate sodium bicarbonate and saline solution; 118250 is dried and fumigated. The crude product Lag pu was crystallized and could be used without further purification. Result: dla), YE (90 7). to

Y 9 —_ _ 'H NMR (400 MHz, CDCI) 8 7.24 (m, 2H), 4.81 (s, 2H), 2.10 (broad, 1 H) (iv) £ cyano = TY - difluorobenzylmethane sulfonate to an ice-cooled solution of ; 1 - difluoro -; - hydroxymethylbenzonitrile (0.7 4 g; 7 mL buffer see step (ii) above) and methanesulfonyl chloride (¥), 1 g; AN 0 mmol) in 01 mL of methylene chloride added triethylamine (AY) + g; AY mmol) with 0 stir after ¥ hours at 0°C; Then the mixture, Cd ye, was washed with 1 M of HCI and once with water + 1187507 and dried and evaporated. The product can be used without further conditioning; Output: 'H NMR (300 MHz, CDCI;) 8 7.29 (m, 2H), 5.33 (s, 2H), 3.07 (s, 3H) (897) p 1.61 (V) 0 4 - Azido methyl-NY-difluorobenzonitrile mixture of 4-cyano = 1,7-difluorobenzylmethanesulfonate) 1.1 g; ) 1.0 mmol; See step iv above) and add sodium 01.0011 ala VY (mol) in 10 mL of water and 00 mL of DMF with stirring at room temperature overnight. The resulting substance was then poured over 701 mL of water and extracted three times with diethyl ether. 1 The combined etherial phase was washed five times with water; And dried with sodium sulfate with fumigation. A simple die was evaporated for NMR purposes and product crystallization. The previous quantity was evaporated with caution, but not to complete dryness. The yield (theoretically 1.77 grams) was assumed to be cumulative based on the analytical HPLC ys NMR.

- © 'H NMR (400 MHz, CDCI) 8 7.29 (m, 2H), 4.46 (s, 2H) (vi); - aminomethyl - 7; +- Difluorobenzonitrile This reaction was carried out according to the procedure used in 3128 (1992) J.

Chem.

Res. (M) To a suspension of OF 10 mg of #1 palladium/carbon (8#1 moisture) in 01 mL of water was added 0 sodium borohydride solution (sodium borohydride) ¢ 0 AY g; ve 71 mol) in Je XY of water. This let out some of the gas. ; -azidomethyl = 6-7-difluorobenzonitrile (VY) g; 4 mmol’ (see step v) (previous) was then dissolved in 0 mL of THF and added to the water mixture on an ice bath for 115 minutes. The mixture was stirred for; hours; to which 10 mL of ? titer of HCL and the mixture was filtered with silite. 0 Celite was rinsed with more water and the combined aqueous phase was washed with EtOAc and then made alkaline with 7 M NaOH. Extraction was performed three times with methylene chloride and then Gut the combined organic phase with water; Drying it on sodium sulfate and evaporation to produce AY (gram A) 7). NMR (400 MHz, CDCI) 8 7.20 (m, 2H), 3.96 (s, 2H), 1.51 (broad, 2H) k 1 1 (vii) ve - difluoro - difluoro - ; - + - butoxycarbonylaminomethylbenzonitrile solution of 4-gid methyl-7; 1-Difluorobenzonitrile (AVI) gram; 5.71 mmol ‘see step (vi) above) was dissolved in 0 mL of THF and di-1-butyldicarbonate (1.5 g; 7" mmol) in 0 mL of THF and then stirred the mixture for 0,000 Y

vy —_ — hour. then evaporate the THF and fractionate the residue between water 5 EtOAc and then wash the organic layer three times with 1.59 M HCL and water; 1482507 dried and fumigated. The product has been used without further purification. The result was 1.748 grams (29 7) (m,2H), 4.95 (broad, 1H), 4.43 (broad, 2H), 7.21 § 1lots 1H NMR (300 MHz, (s, 9H) ° 1.52 Boc-Pab(2,6-diF)(OH) (viii) mixture of 7 + - difluoro = t = -butoxycarbonylaminomethylbenzonitrile VA) g; 5.16 mmol; see step (previous vid); Hydroxyamine hydrochloride 10155 cal ja 0.1 (mmol) and triethylamine 0.57 (g” 100 mmol) were stirred in 01 deg ethanol with stirring at room for 37 hours. The solvent was evaporated and the residue fractionated between water and methylene chloride. The organic layer was washed with water, dried and evaporated. The product can be used without further purification. The result is 0.47 grams (97 7). 'H NMR (500 MHz, CD;0D) 5 7.14 (m, 2H), 4.97 (broad, 1H), 4.84 (broad, 2H), (broad, 2H), 1.43 (s, 9H) 4.40 Boc-Pab(2,6-diF) x HOAc (ix) 0 This reaction was carried out according to the procedure described in: V,YY) Judkins etal, Synth.

Comm. (1998) 4351. Boc-Pab(2,6-diF)(OH) girly £,YV mmol; See step (viii) above); then acetic anhydride (vv) 0 g was added; Excelled

vy _— _ mmol) and 7 ;; mg Gel)ye palladium/carbon (H 75 0) in 100 mL acetic acid was hydrogenated at © atmospheric pressure for 3.5 hours. The mixture was filtered through celite, rinsed with ethanol, and evaporated. The residue was lyophilized from acetonitrile, water, and a few drops of ethanol. The resulting subtitle complex can be applied to acetonitrile©, water and a few drops of ethanol. This compound can be used without further purification; Output 4 g (749) (m, 2H), 4.34 (s, 2H), 1.90 (s,3H), 1.40 (s, 9H) 7.45 8 sq'H NMR (400 MHz, Boc-Pab) (2,6-diF)(Teoc) (x) to a solution of HOAC Boc-Pab(2,6-diF) ) 1.071 g; 5.44 mmol; see step (ix) 0 preceding) in 100 mL of THF and 1 mL of water were added 7-(trimethylsilyl)ethyl m-nitrophenyl carbonate 1.1V) g; 5.84 mmol). and a solution of potassium carbonate (1.07 g + eV mol) in yo ml water was added dropwise over © minutes. Then the mixture was stirred overnight. The THF was evaporated and the residue fractionated between water and methylene chloride. The aqueous layer was extracted with methylene chloride and the combined organic phase was washed twice with aqueous VO sodium bicarbonate; And drying it on 1182507 and fumigating it. Flash chromatography on silica gel with heptane/20/6 = 7/1 yielded 1.11 g (VY 7) of the crude compound. '"H NMR (400 MHz, CDCI) 8 7.43 (m, 2H), 4.97 (broad, 1H), 4.41 (broad, 2H), (m, 2H), 1.41 (s, 9H), 1.11 ( m, 2H), 0.06 (s, 9H) 4.24 \Voy

—-YY-

Boc-Aze-Pab(2,6-diF)(Teoc) (XI) prev) X g; 7.76 mmol; See step 0001 4) Boc-Pab(2,6-diF)(Teoc) minutes; And evaporate it 0 then leave the mixture for a while. (#2) HCl — saturated 0 mL EtOAc dissolved in ; g;

gram; 7,725 mmol) and at the end diisopropyl (V,YE) PyBOP mmol) 7,714 hours and then sad was added. The reaction mixture was stirred (Use ml AAT g; 1.1 0A) amine. The organic phase EtOAc ml water was washed and extracted three times with YO pour over and evaporated. Flash chromatography on a gel (NaySO4) collected in brine; drying it with silica with heptane 1 EtOAc: : 9 yielded 0.091 g (427) of the desired compound. (m, 2H), 4.65-4.5 (m, 3H), 4.23 ) 2H), 3.87 1 7.46 8 (total 1H NMR (500 MHz, (m, 1H), 3.74 (m, 1H), 2.45-2.3 (m, 2H), 1.40 ),911(, 1.10 (m,2H), 0.05 (s, 9H) Ph(3-CI)(5-QCHFp)-(ff) CH(OH)C(0)-Aze-Pab(2, 6-diF)(Teoc) (xii) Boc-Aze-Pab(2,6-diF)(Teoc) ) 01 dissolved 7 g; 1,950 mmol; see step (xi) above) in 1 mL of EtOAc saturated with HCL (g). the mixture was left for 10 minutes; 1 was then evaporated and dissolved in ©mL of Ph(3-CI) (5-OCHF)-(fl)CH(OH)C(O)OH .DMF 0 ).g; 5 ml of filler (see Preparation (viii) A above); 1 ¥) PyBOP grams; 9A, + mmol) and finally diisopropylethylamine (£0 Y, + g” 1.84 mmol) was added. The reaction mixture was stirred for 2 hours, then it was poured over water and extracted three times with a combined organic shall EtOAc, washed with brine and dried thoroughly.

- vs — Sodium sulfate and fumigation. Flash chromatography on silica gel with EtOAc yielded 1,184 grams (770) of the compound contained in the desired subtitle. 'H NMR (400 MHz, CD3OD, mixture of rotamers) 8 7.55-7.45 (m, 2H), 7.32 (m, 1H, major rotamer), 7.27 (m, 1H, minor rotamer), 7.2-7.1 (m, 2H ), 6.90 ) 1H, major rotamer), 6.86 (t, 1H, minor rotamer), 5.15 (s, 1H, major rotamer), 5.12 (m, 1H, ee minor rotamer), 5.06 (s, 1H, minor rotamer), 4.72 (m, 1H, major rotamer), 4.6-4.45 (m, 2H), 4.30 (m, 1H. major rotamer), 4.24 (m, 2H), 4.13 (m, 1H, major rotamer), 4.04 (m, 1H, minor rotamer), 3.95 (m, 1H. minor rotamer), 2.62 (m, 1H, minor rotamer), 2.48 (m, 1H, major rotamer), 2.22 (m, 1H, major rotamer), 2.10 (m, 1H, minor rotamer), 1.07 (m, 2H), 0.07 (m, 9H)

Ph(3-CI)(5-OCHF;)-(ff)CH(OH)C(0O)-Aze-Pab(2,6-diF)(OMe,Teoc) (xiii) a mixture of Ph(3- CI)(5-OCHF,)-(H)CH(OH)C(O)-Aze-Pab(2,6-diF)(Teoc) ) 1 mg 4 mmol; see step (xi) above) and 0-methylhydroxylamine hydrochloride (04 mol 1.20 mmol) in ; mL of acetonitrile heated at Ve °C for 1 ? hours. The solvent was evaporated and the residue was fractionated between water and (2)0. The aqueous layer was extracted twice with B-BtOA and the organic phase was washed with water, dried with sodium sulfate, and evaporated. The product can be used without further purification. Output 0A mg AY (7).

__- Yo — 'HNMR(400MHz, CDCI3) 5 7.90 (bt, 1H), 7.46 (m, 1H), 7.25-6.95 (m, SH), 6.51, t, 1H),4.88(s, 1H),4.83 (m, 1H), 4.6-4.5 (m, 2H), 4.4-3.9 (m, 4H), 3.95 (s, 3H), 3.63 (m, 1H), 2.67 (m, 1H), 2.38 (m, 1 H), 1.87 (broad, 1H), 0.98 (m, 2H), 0.01, s,9H)

Ph(3-CI)(5-OCHFp)-(ff) CH(OH)C(0)-(S)Aze-Pab(2,6-diF)(OMe) (xiv) oA) Ph(3-CI )(5-OCHF,)-(fl)CH(OH)C(0)-Aze-Pab(2,6-diF)(OMe,Teoc) 0 was dissolved with cooling on TFA pre) in ¥ ml of (xiii) mg; 7, _ ml filler, see step, and dissolve the remaining material in 110/86. TFA was ice bathed and allowed to react for 2 hours. (Na2S04) The organic layer was washed twice with aqueous sodium carbonate and water, dried at (79%) mg £Y and evaporated. The residue was evaporated from water and acetonitrile to provide 0.0 of the compound given in the title. 'HNMR (300 MHz.CDCI3)S 7.95 (bt, 1H), 7.2-7.1 (m, 4H), 6.99 (m, 1H),6.52(t, 1H), 4.88(s, 1 H), -4.85 4.75 (m, 3H), 25 4.6-4.45 (m, 2H), 4.29 (broad, 1H), 4.09 (m, 1H), 3.89 (s, 3H). 3.69 (m, 1H), 2.64 (m, 1H), 2.38 (m, 1H), 1.85 (broad, 1H)

BC-NMR (100 MHz; CDCI): (carbonyl and/oramidinecarbons) 5172.1,169.8,151.9

APCI-MS: (M + 1) = 533/535 m/z Vo \VoY

Preparation of compound E (Ph(3-CI)(5-OCH,CHSF)-(R)CH(OH)C(0)-(S)Aze-Pab(OMe)) V)(i) mono Fluoroethyl) methane sulfonate to a solution that is magnetically stirred from? - fluoroethanol (0,0 g); 78650 mmol) in (11012 (Je 01) in a nitrogen atmosphere at 0 °C were added triethylamine het (Y vv) g; ? Je mol) and methanesulfonyl chloride (Yo VY) g; 1 mmol). Then the mixture was stirred at 0 °C a h , 1 h and diluted with (Se 1 00 ) CH,CI, and washed with Y Uke of (001 HCl ml). The aqueous layer was extracted with (de 04) CHCL and the organic layer was extracted, washed with brine (JoVO), dried with sodium sulfate, filtered, and concentrated in a vacuum to afford the compound given in the subtitle (no q, g g 0 ). 0 A yellow oily substance that was used without further purification. = Hz, 1 H), 4.43 (t, J= 4 Hz, 1H), 4 (s, 3H). (if) ¥ chloro — 0 — 1 monofluoroethoxybenzaldehyde into solution from ? -chloro-0-hydroxybenzaldehyde (AY) g” 0Y,0 mmol” see previous preparation (i)A) Potassium carbonate (9.4 g; TAY mmol) was added in ( Je (V+) DMF in a nitrogen atmosphere to which a solution of (?-monofluoroethyl)methane 4,V) g was added; TAY mmol See step (1) above) in a (171 mL) DMF drip at room temperature. The reaction mixture was heated to 100 °C for several hours

Ala - then stirred overnight at room temperature. The reaction was cooled to zero degrees Asie and poured over ice-cold HC? Titer and extract with EtOAc. The collected organic layer extracts were washed with brine; dried with sodium sulfate; And filter and focus in a space of space. Brown oil was treated by chromatography on silica gel by eluting with (1 : £) EtOAc : Hex to provide the compound given in subheading (6," g; VY #) as oily yellow. NMR (300 MHz, CDCI3) 6 9.92 (s, 1H), 7.48 (s, 1H), 7.32 (s, 1H), 7.21 (s, 111 50 1H), 4.87 (t, J= 4 Hz, 1H), 4.71 (t , 123 Hz, 1H),4.33(t, J=3Hz, 1H), 4.24 (t, J= 3 Hz, 1H). Ph(3-CI)(5-OCHCH:F)-(fl 0,S)CH(OTMS)CN(LI) to a solution of *-chloro-0-monofluoroethoxybenzaldehyde (cal ja V1) 7.5 mmol; see step (n) above and zinc iodide (clon Te) 9.78 mmol) in 112012©) + (Jo™Y) trimethylsilyl cyanide (4," g; 75.0 mmol) was added dropwise at 0 °C Asha in a nitrogen atmosphere. The reaction mixture was stirred at 0 °C for 1 h at room temperature overnight The reaction mixture was diluted with water (300 mL), the organic layer was separated, dried with sodium sulfate, filtered, and concentrated in space to provide the compound in the title sub (VT) grams; 94#) as a brown colored oil that has been used without further purification or diagnosis. come back

YA — _ Ph(3-CI)(5-OCH,CH-F)-(f1,S)CH(OH)C(O)OH (iv) concentrated hydrochloric acid (001 mL) added ) to : 0A ala 0 ,( Ph(3-CI)(5-OCH,CH,F)-(1,S)CH(OTMS)CN mMol see step (iii) previous) and the solution made Stir it at 100°C for ? hours. After © cooling to room temperature; The reaction was cooled further to 0 °C; He slowly acidified it with 7N of NaOH - (Je Yeo) and washed it with (Ja Yoo X ¥) Et,O. Then acidified the hydrated layer with ?N of 1101 (Je A+) and extracted it with X ¥ ) EtOAc (Sa Yeo). The (Na;S04) EtOAc extract was then dried, filtered, and concentrated in vacuo to provide the compound in heading AT) grams; 98 7) as a yellow solid was used without 0 - further purification. Ft, = 0.28 (90:8:2 CHCI3:MeOH:concentrated NH,OH) 'HNMR(300MHz, CD;0D) 5 7.09 (s. 1H), 7.02 (s, 1H), 6.93 (s, 1H), 5.11 (s, 1H), (m, 1H), 4.62-4.65 (m, 4.77-4.81 1H), 4.25 -4.28 (m, 1H), 4.15-4.18 (m, 1H). (Vv) \o : Ph(3-CI)(5-OCHpCHpF)-(S)CH(OACc)C(O) OH (a) and Ph(3-CI)(5-OCH,CHpF)- (f)CH(OH)C(O)OH (b) dla

A solution of 0 4,6 « al ya A,1) Ph(3-CI)(5-OCH,CH,F)-(R,S)CH(OH)C(O)OH mmol was heated; See step Lipase PS "Amano" ys «(iv) (1.; g) in vinyl acetate You) (Je YOO) MTBEj (Je) heated at Ve °C in nitrogen for Days. The reaction mixture was cooled to room temperature and the enzyme was removed by filtration through celite. M filter paste was washed with B 200/806 and concentrated in a vacuum space. After filtration using chromatography on silica gel B EN: MeOH: CHCl; (0 : + : ) to provide the triethylamine salt of the compound given in subheading (a) as yellow oil.In addition, the triethylamine salt of the compound given in subheading (b) was obtained (1,;; g). It was acidified with 0 ¥ mL of HCL and extracted with Yoo X ¥ (EtOAc mL). The combined organic extract was dried on 08250; was filtered and concentrated in space to yield the compound given in subheading YA) (b) grams; FY 7#) as a yellow oil. Subtitle compound data (b) R= 0.28 (90:8:2 CHCI3:MeOH:concentrated NH,OH) 'H NMR (300 MHz, CD;0D) 5 7.09 (s, 1H), 7.02 (s, 1H), 6.93 (s, 1H), 5.11 (s, 1H), Vo (m, 1H), 4.62-4.65 (m, 30 1H), 4.25-4.28 (m, 1H), -4.15 4.18 (m, 1H). a solution of cane AYA) Ph(3-CI)(5-OCH,CH,F)-(R)CH(OH)C(O)OH 7.79 mmol; See previous step v) in Ye (DMF ml) in a nitrogen atmosphere at 0 °C for

4. £,YV cal 2 (0.47 HCI 1 HAze-Pab(OMe) mmol) added; see ip.application 42059/00 (WO 27307 (845, g » 7.18 mmol); 5 0.1 1 (DIPEA 1 g; ALYY mmol). The reaction was stirred at 0 °C for 2 hours and then at room temperature overnight. The mixture was concentrated in a vacuum and a chromatography was performed twice for Substance 0 remaining with silica gel and then filtered first with 01101: (1 : 10) EtOH and then with EtOH : EtOAc (VY) to provide the compound given in heading (cama ANY) 54 7). CHCI;:EtOH) 'H NMR (300 MHz, CD;0D, complex mixture of 10:1( 0.60 grain rotamers) 8 7.58-7.60 (d, J= 8 Hz. 2H), 7.34 (d, J = 7 Hz, 2H), 7.05-7.08 (m, 2H), -6.95 (m, 1H), 5.08-5.13 (m, 1H), 4.77-4.82 (m, 1H), 4.60-4.68 (m, 1H), 3.99-4.51 (m, 6.99 7H), 3.82 (s, 3H), 2.10-2.75 (m,2H).*C-NMR (150 MHz; CD;OD): (carbonyl and/or 00 Amidine carbons) 5 173.3, 170.8, 152.5. APCI-MS: (M + 1) = 493 m/z. Preparation of compound F (Ph(3-CI)(5-OCHFo)-(ff)CH( OH)C(0)-Aze-Pab(OH)) (i) Ph(3-CI)(5-OCHFp)-(fl)CH(OH)C(O)-Aze-Pab(OH, Teoc ) dissolved 074 cal ya +) £¥) Ph(3-CI)(5-OCHF;)-(fl)CH(OH)C(0)-Aze-Pab(Teoc) \o mL mol' see. Prepare step (ix) D above) in mL of acetonitrile and 0.10 g V0 (mmol) of hydroxylamine hydrochloride. The mixture was heated at Ve °C for 7.5 hours; Then it was filtered through the silite and evaporated. Crude product was used (£0 101 g; VO purity) directly into the next step without further purification). Dla

Ph(3-CI)(5-OCHF,)-(ff) CH(OH)C(O)-Aze-Pab(OH) (ii) g;VE°) Ph(3-CI)(-5 OCHF,)-(fi)CH(OH)C(O)-Aze-Pab(OH, Teoc) 43 TFA and 9 mL of the former CHoCly) were metabolized in 50 + mL of (i) See step edge Jao, YY

HPLC and the remaining 480 were done with TFA. Evaporated 0 min Tv allowed to complete the reaction for 100 mg (yield on preparative H/Al. Then the significant particles were decanted and lyophilized (twice); to produce two steps #77 of the compound mentioned in the title.‎

MS (m/z) 482 (M - 1)-; 484 (M + 1)+' H-NMR (400 MHz; CD; 0D): 8 7.58 (d, 2H), 7.33 (m, 3H), 7.15 (m, 2H), 6.89 (t, 1H major rotamer) , 6.86 ) 1H minor 5 rotamer), 5.18 (s, 1H major rotamer; andm, 1H minor rotamer), 5.12 (s, 1H minor rotamer), 4.77 (m, 1H major rotamer). 4.42 (m, 0 2H). 4.34 (m, 1H major rotamer). 4.14 (m, 1H major rotamer), 4.06 (m, 1H minor rotamer), 3.95 (m, 1H minor rotamer), 2.66 (m, 1H minor rotamer), 2.50 (m, 1H major rotamer). 2.27 (m, 1H major rotamer), 2.14 (m, 1H minor rotamer) BC-NMR (100 MHz; CD3;0OD): (carbonyl and/or amidine carbons, rotamers) 5 172.4, 172.3, 172.0, 171.4 152.3, 152.1 Vo

H (Ph(3-CI)(5-OCHpCHFp)-(ff)CH(OH)C(O)-Aze-Pab(OH)) Compound preparation

Ph(3-CI)(5-OCH.CHFp)-(fl)CH(OH)C(O)-Aze-Pab(Z) (i) dla

_ Y _ Boc-Aze-Pab(Z) (see alla INP No. 07744 [ AY « AY mg; 6.197 mmol] was dissolved in 01 mL of 6/260 saturated with HCI (grams) and allow the reaction to take — minutes. Then evaporate the solvent and then mix the remaining 0

PyBOP mmol); 164 mg 0 ) Ph(3-CI)(5-OCH,CHF,)-(fi)CH(OH)C(O)OH mmol) and eventually diisopropylethyl amine) 47 mg; 78 mL 17094 mg; (144 0 mL of 09) then the mixture was stirred for 2 hours, then poured over 5 mL of water (Y mol) in the combined organic phase, washed with water, dried, EtOAc, and extracted three times b Then, the flash chromatography of the crude product was done on silica gel using 0 ad 9 (B212250) (LAY) 001 mg yield (1): MeOH: EtOAc 'H NMR (300 MHz, kum mixture of rotamers) 5 7.85-7.75 (m, 2H), 7.45-7.25 (l7) 0 7H), 7.11 (m, 1 H, major rotamer), 7.08 (m, 1H, minor rotamer), 7.05 -6.9 (m, 2H), 6.13 (bt, 1H), 5.25-5.05 (m, 3H), 4.77 (m, 1H, partially hidden by the CD3;OH signal), 4.5-3.9 (m, 7H), 2.64 (m, 1H, minor rotamer), 2.47 (m, 1H, major rotamer), 2.25 (m, 1H, major rotamer), 2.13 (m, 1H, minor rotamer) \o Ph(3-CI)(5- OCHpCHF,)-(ff)CH(OH)C(0O)-Aze-Pab(OH) (ii)

VA) hydroxylamine hydrochloride (10 mg) was mixed; 94 mmol) and triethylamine 60 and treated with ultrasound for one hour at THF ml of A mmol at 3.17 cal a ( Ph(3-CI)(5-OCH,CHF,(-) fl)CH(OH)C(0)-Aze-Pab(Z) °C added after

_ $ y _ 401 Then the mixture was stirred at .THF ml of A (previous) with (i) ml mole see step 1, 0 mg preparative with RPLC °C for 6 days . The solvent was evaporated and the crude product b was purified

Jaa Purity: (7 YA) mg Ye Yield: (1 6:06 ) NH;O0Ac molar of 1,1 : CH;CN '"H NMR (300 MHz, CD;0D. mixture of rotamers) 8 7.6-7.55 (m, 2H), 7.35-7.3 (m, 2H), 7.12 (m, 1H, major rotamer), 35 7.09 (m, 1H, minor rotamer), 7.05-6.9 (m, ° 2H), 6.15 (triplet of multiplets, 1H), 5.15 (m, 1H, minor rotamer), 5.13 (s, 1H, major rotamer), 5.08 (s, 1H, minor rotamer), 4.77 (m, 1H, major rotamer ), 4.5-4.2 (m, 5H), 4.08 (m, 1H, major rotamer), 3.97 (m, 1H, minor rotamer), 2.66 (m. 1H, minor rotamer), 2.50 (m, 1H major rotamer), 2.27 (m, 1H, major rotamer), 01 2.14 (m, 1H, minor rotamer).

BC-NMR (100 MHz; CD3; OD): (carbonyl and/or amidine carbons, mixture of rotamers) 8 172.8, 172.2, 171.4, 159.1, 40 158.9,154.2.

APCI-MS: (M + 1) = 497/499 m/z

YoY

- oss Abbreviations acetyl Ac (MS) atmospheric ionization (related to APCI) MS atmospheric ionization (related to API aqueous aq ° carboxylate (unless otherwise specified) - 1 - Azetidine — (S) Aze & (S)-Aze butyloxycarbonyl - 1 Boc broad (in relation to NMR) br (MS in relation to La) chemical ionization cl dd(s) ) d 01 bi (for NMR) d dicyclohexylcarbodiimide (DCC) for NMR (Lad) bifold dd diisobutyl aluminum hydride (DIBAL-H) Isopropylethylamine z DIPEA \o \Yoy

DH - NON) - ¢ DMAP - Dimethylamino)pyridine DMF 7 Dimethylformamide DMSO Dimethylsulfoxide 050 Differential Scanning Calorimeter DVT |- 0 Internal Vein Thrombosis 1 EDC - ( © - dimethylaminopropyl) - © - ethyl carbodiimide hydrochloride eq.ES equivalents electrospray ESI electrospray intermediate Et 0 ethyl ether diethyl ether EtOAc ethyl acetate EtOH Ethanol Et,0 m diethyl ether HATU 0 - (azabane zotriazole = 1 - yl) - NNNON' - tetramethyl Voy

uranium hexafluorophosphate HBTU 1,11-tetramethyl-©-(benzotriazole-1-yl)z uranium hexafluorophosphate HCI HCl; Hydrogen chloride gas or Salt ° Hydrochloride (depending on context) Hex Hexanes HOAc Acetic acid HPLC High-performance liquid chromatography LC Liquid chromatography d Multi-Lad (NMR related) Me Methyl MeOH Methanol min.m(s) MS Mass Spectrum dont Jus MTBE \o Ether YVo¥

gv - - NMR OAc NMR Acetate Z Pab Para-Amidinobenzylamino H-Pab Para-Amidinobenzylamine Pd/C° Palladium on Ph-Carbon Phenyl PyBOP (benzo Triazole-:-yl-oxy)tripyrrolidinosphosphonium hexafluorophosphate q quaternary (for NMR) 00"0 tetrabutylammonium fluoride 181 room temperature 8 single (for NMR) triple (for NMR) NMM) NNN'NJ TBTU - tetramethyl - © - (benzotriazole-1-yl) \o uronium tetrafluoroborate] \voY

— $A —_

TEA triethylamine

‎Teoc ¥ - (trimethylsilyl)ethoxycarbonyl

TY oY TEMPO - tetramethyl = 1 - piperidinyloxy without

TFA moieties are trifluoroacetic acid

TGA is a thermogravimetric analysis

THF tetrahydrofuran

TLC thin layer chromatography

ultraviolet radiation

The prefixes 0, 5, and 1 have their traditional meaning: normal, secondary, iso, and tertiary. Examples VE - Ago - 1 illustrate the present invention. Examples 76 and each are provided for comparative purposes only and do not form part of the present invention. In the examples and figures the ratios given in parentheses refer to the weight ratio of the neutral gel polymer to iota-carrageenan and the base pharmaceutically active ingredient or any other ingredient that may be present is not taken into account. Answer

q — _ Brief explanation of the drawings in the accompanying figures: Figure 1: Release of H376/95 from mixtures with different synthetic ratios of PEO-iota-carrageenan; Molar. The tablets were analyzed for 2 hours at 1 pH and for the specified time at 0 LA pH Figure 7: Release of H376/95 from mixtures with different compositional ratios (A+: 71) from 7120 with Different molecular weight ratio and Iota-carrageenan The tablets were analyzed for two hours at a pH of 1 pH and for the specified time at a pH of 11m SUA Figure ?: Release H376/95 from mixtures with different compositional ratios (01: A+ ) of 17120 with 0 having different molecular weight ratio and Iota-carrageenan. The tablets were analyzed for two hours at a pH of 1 pH and for the specified time at a pH of UA PH Figure: release of H376/95 from mixtures with different synthetic ratios of Iota-carrageenan and (HPMC) 0 cP. Analysis of the tablets for two hours at pH 1 pH and for the specified time at pH > 1 A pH , 3 vo Fig. 10 Release of H376/95 from mixtures with synthetic ratios (04 : +0) PEO; Molar"; and Iota-carrageenan. Tablets were analyzed for YE 1 h in different media. Figure 1 Release of H376/95 from a neutral PEO gel polymer; Molar analyzed in different g 3 culture media. \Voy ‏

- and ©_—

Figure 7: Release of H376/95 from anionic polymer Iota-carrageenan analyzed in industrial media

3 § different.

Figure 8: Release of compound A from mixtures with molar ratios (00:00) of Iota-carrageenan and

PEO; Molar at a pH of 1, pH 1.8. The tablets were analyzed for YE hours in 0 different media.

Figure 24 Release of compound A from mixtures in molar ratios (0 9 : 00) of Iota-carrageenan and

00001 HPMC centipeas, pH 1, pH 8,. has been analyzed

the tablets for YE hours in different industrial media.

Figure 01: Release of Complex B from Iota-carrageenan mixed with the neutral polymer PEO; Molar (0) in a ratio of 50 pon) and (80: 01). The tablets were analyzed for two hours at a pH of 1 pH.

and residence time at LA pH

Figure 11: Release of compound 3 of Iota-carrageenan mixed with neutral polymer (HPMC)

0 centipeas at a ratio of (00:20). The discs were analyzed for 2 hours at pH

LA pH and residence time at 1 pH

Dla

1 _ M Cia ol Detailed Example 1 This example shows the release of H376/95 from mixtures with different compositional proportions of PEO; Molar and Iota-Carrageenan. The tablets were analyzed sad 2 hours at a pH of 1 pH and for a period of time at SLA percentage PEO; Molar: (As YY) (04:04) (Ye: AY) - sd carrageenan > y oH | 2 The tablets were made by direct pressing The active ingredient of PEO was mixed; Molaro Lisl Carrageenan. completely with 0 lubricant and then sodium stearyl fumarate added » through a sieve of 0,000 mm. Further final mixing was performed and the mixture was pressed using an additional 9 mm bore in a single hole 0 tablet press. The tablets were analyzed in a bath Melting; UPS 11 (04 rpm at 317 °C; synthetic medium) containing 11 M HCL at 1 pH for 2 hours. The tablets were then moved to a bath Dissolved with 11 M phosphate buffer, pH 1.8, and analyzed again A. Display results in \Voy

Y —_— 0 - Figure 1. The formulation (00 : 00) showed an initially pH-independent release form at a pH between LA - 1. Bad can be concluded that at Mixing different ratios of anionic polymer Iota-carrageenan and neutral gel polymer PEO Molar The rate of release into the medium can be adjusted by using a different pH °Example ¥: This example showed that the release of a H376/95 mixtures with a ratio Composition (A+ : Yo) of PEO with different molar ratios and Iota-carrageenan The tablets were analyzed for two hours at pH 1 pH and for the remaining time at pH SLA Co polyoxide 0 ltd mg Polyethylene oxide Sodium stearyl fumarate 8 mg Sodium stearyl fumarate 0 Tablets were manufactured and analyzed according to example 1. The results of the analysis are shown in Figure ¥ which shows that using a higher molecular weight of the neutral gel polymer results in a higher release rate lower in dl number

Y _ m neutral pH. The release rate is not affected in the lower pH region due to the presence of sufficient amount of anionic polymer in the formulation. l » l y This example shows the release of 11376/95_ from mixtures of composition ratios Av(:70) of 171250 with 0 different molecular weight to iota-carrageenan. The tablets were analyzed for two hours at a pH of 1 pH and the remaining time was at a pH of SLA pH, then the tablets were prepared and analyzed according to example 0 1 Figure 1 shows how using a gel polymer with a high molar weight can reduce Release rate at neutral pH. 0 At the same time the release inactivation rate at pH 1 has less discreteness due to the use of less Iota liad HIS compared to the examples shown in XSAN \Voy

— M Example 4 This example shows the release of 11376/95 from mixtures with different synthetic ratios of iota-carrageenan and 110140' 00001cP. The tablets were analyzed for 2 hours at 1 pH and the rest time at LA pH o THPMC ratio Iota - (As :Y4) (0+:04) (Y+ :AY) Carrageenan

101 The tablets were manufactured and analyzed according to example 1. The results of the analysis in a different melting medium are shown in the figure Lg The formula (04 : 00) again showed a non-primarily pH-dependent release at a pH of 011 ranging from UA - 1 It can be concluded again that the release rate can be modified by mixing a range of ratios of neutralizing gel polymers;

ye in this case 00001 HPMC centipoise; With the anionic polymer Gad Carrageenan.

o Jud; Molar and PEO of ( Cv: On ) This example shows the release of 5 ratio mixtures of Iota-carrageenan formulation. The discs were analyzed for; 7 hours in different media Molar: Eddie § PEO Ratio Iota - Carrageenan: Noli - | Molar > The analyzes were carried out in baths 1. The tablets were manufactured by direct pressure according to Example 0 with the tablets in a container on the flow stream) where three tablets are analyzed (Y USP lea) Molar melting of the buffer solution 1,1 phosphate and HCI molar of 1 Jag in each aclu for a period of ; ? is added to improve the potential (EtOH) with © #ethanol A pH molar with pH 1 showing that the tablet with the release of the co-solubility of the drug. The results in the figure

pH-dependent PEO can be made; when using a composition having equal parts of 0 m basket made of quadrangular wire interwoven; meat from TT; Molar and Iota-carrageenan. One of the small upper flanks to the end of the solid cord. The rope is made through a cover cm from the center of the vessel. The YY end is the “melting pan” and is secured with two Teflon \Voy knots

A — m — the lowest of the special bottle is adjusted so that it is 1 cm from the stirrer. The inrush stream is directed with the wail under test at its edge.] . Ju 3 1” This example shows release of H376/95 from PEO neutral gel copolymer; Molar in the absence of Iota-0-carrageenan and its analysis in different synthetic media.

Het od polyoxide rd er

The tablets were made by direct compression according to Example 1. The analyzes were carried out separately in different melt baths. The tablets in vessels containing Ge Ve 1.1 HCI were analyzed for two hours 0. when using a buffer solution of 0.11 M phosphate ¢ with a pH

0 1,8 as dissolving media; The tablets were analyzed for 71 hours. The results in Figure 6 are shown; that

The release rate at 1 pH is significantly greater than the release rate at

pH CLA indicating that the use of a neutralizing polymer is not sufficient to provide a non-CLA release

The pH-dependent of a major drug with pH-dependent solubility. Aaa da

Y —_ 0 —— EXAMPLE 1 This example shows the release of H376/95 from the anionic polymer iota-carrageenan in the absence of a neutral gel polymer and its analysis in different media. Under sodium stearyl ionomer 0 the tablets were made by direct pressing according to Example 1. The analyzes were carried out separately in different solubilization baths; Similar to Example 7. Figure 1 shows how the release rate at pH 1 is lower compared to release at pH 6.8. This effect is not seen when any of the other homopolymers we tested are used as a group polymer. Example A 0 This example shows the release of compound A from a mixture with a formula ratio (50 : 00) of PEO, Molar and Iota-carrageenan. Yad tablets were analyzed 1 hour in different media. , A Iota-Carrageenan Vou, (Fluka) PEO, 4M (Union Carbide) M1 PRUVT™ Y,0 The active ingredient was manually mixed with polymers and lubricants.The mixture was pressed directly into tablets l

A — © Example 9 This example shows the release of compound A from a mixture with a formula ratio (00 : #0 ) of 00001 HPMC centi-Poise and Iota-carrageenan. The YE add discs were analyzed 1 hour in different media. dod Compound On ,* A Iota - Carrageenan Ver, (Fluka) 0 HPMC CPT 011 Yo PRUVT™ 0 The active ingredient has been hand-blended with polymers and lubricants. The mixture was pressed directly into tablets. Cumulative Release Evaluation of A Spal) from the tablets in Examples A and 1 Two tablets were tested for drug release in 900 mL media using a USP ? (Stickers + Basket 1) at 00 rpm at TV°C. The dissolving media used were 1.1 M of hydrochloric acid (pH 1) and 11 M of a buffered solution of Yee sodium phosphate (LA pH) with ©7#ethanol added to improve The ability to dissolve the drug The addition of ethanol was different so as not to affect the rate of release of these components significantly Consistent accumulation was performed using a fiber-optic vomit © system with an analytical wavelength of 7705 nano jie when using 1.1 M HCL As a melting medium with an analytical wavelength of nanometers when using a modified buffer solution of phosphates with a pH and

_ 9 M 1A as a melting medium. Then use the length of Yo. ds ga nm as reference length in each of Example 01 This example shows the release of compound B from a mixture with a composition ratio of 8+0 : 860 of PEO; Caliber and 0 Iyota Cargenan. J bq al mq is The tablets were made according to Example 9. The release data is shown in Figure 01. Example 11 This example shows the release of compound 33 from a mixture with a composition percentage (A) of PEO Molaro 0 Iota Carrageenan DMLA

= .Te TT composite cradle 8 e Basket made quadrangularly of intertwined wire; It was welded from one of the small upper sides to the end of the steel rope. The rope is threaded through the lid of the melting pot and secured with two Teflon knots” VY cm from the center of the pot. The lower end of the special bottle is 0 adjusted so that it is at a distance of 1 cm from the stirrer. The effluent stream is directed with the disc under test at its edge]. The tablets were made according to Example 9. The release data are shown in Figure 01. Example 11 This example shows the release of Compound B from a mixture with a synthetic ratio (+0 : +0) of HPMC 0111 01 Sinte Boise and Iota-Carrageenan.

a 0 0 pm 0 7 ¢ \ \ 0 $ HPMC m ( 111 centipoise ) (SH Te) Discs were made according to Example 4. Release data is shown in Figure AN Example 11 Direct compression of the compound © with 00001 HPMC cP° then mix the active substance and excipients in a large container then lubricate the mixture with sodium stearyl fumarate and compress it into tablets using a press tool Sodium Stearyl Fumarate Y,0 mg IY en Sodium stearyl fumarate No

Y _ 4 -_ release rate data time (min) release rate in solution | The release rate in the pH buffer solution | Regulator pH 1.1 LA AH EL WW OE EE EE TR EE Direct Pressure Compound C B 00001 HPMC Centine Boise and Iota Carrageenan; At a ratio of .5 : 06 ° how much to mix the active substance and the excipients in a large container. The mixture was then lubricated with sodium stearyl fumarate and compressed into tablets using a press. fill

- Sa m EI Sodium stearyl fumarate Release rate data Time (min) Release rate in solution | The rate of release in the buffered solution with a pH 1_ the regulator with a pH LA 11 except for the aaa aath aaath aaaaaaaaaaaaaaaaaahah ee

YoY

—-— 1¢ —_

The release rates were determined as follows. Three individual drug release tablets have been tested in

0 mL of medium using Y USP dissolving device (stirrer + basket 1) at 00 rpm

at VY °C. The dissolving media used were 11 M of hydrochloric acid (pH 1) and 11 M of sodium phosphate buffer (pH

0 pH (VA PH) Consistent accumulation was performed using the © Technologies fiber optic system with an analytical wavelength of 701 nm when using 1.1 M 1101 as melting medium and with an analyte wavelength of 0 nano jie at Use a modified buffer solution of phosphate with a pH

1A as melting medium. Then use the You Nano Be wavelength as a reference length in each of the two mediums. In the first 2 hours of analysis, the release value was measured every VO min; Then every hour rest

,. Tahlia Al waqt 1 01 4 Example A w | ow a oo sodium stearyl fumarate \Voy

_— 0 — This formulation is prepared as given in Example AY A 1 Basket made of four corners of intertwined wire; It was welded from one of the small upper sides to the end of the steel rope. The rope is threaded through the lid of the melt pot and secured with two Teflon loops 3.7 cm from the center of the pot. The lower end of the special bottle 0 | is adjusted so that it is 1 cm from the stirrer. The effluent stream is directed with the disc under test at its edge]. humiliation

Claims (1)

Claims 1 1 - Oral pharmaceutical formulation comprising “gota-carrageenan (52a) or more neutralizing gelling polymers 3 5 and a basic pharmaceutical active substance pharmaceutically active “ingredient ¢ where this formulation inhibits the release of a major pharmaceutical active ingredient from this formulation at an acidic pH 1 . 1 formulation according to claim 3 where the component is The pharmaceutically active has a base group of 1 epKa to .01 1 - composition.formulation according to claim 1 or 1 wherein the pharmaceutically active base Y is ximelagatran or ximelagatran Ph(3-CI)(5-OCHF,)-(R)CH(OH)C(0)~(S)Aze-Pab(OMe); Ph(3-CI)(5-OCHF,)- v (fi)CH(OH)C(O)-(S)Aze-Pab (2.6-diF)(OMe);orPh(3-CI)}(5-OCH,CH,F)- ¢ (R )YCH(OH)C(O)-(S)Aze-Pab(OMe).© 1 — — formulation according to claim 1 where the base component Li all Y is pharmaceutical HEY) 3-dimethyl-2-oxobutyl)-9-oxa-3,7-diazabicyclo [3.3.1] non-3 yl] ¥ ,3( -7[ =3{ (-4 propyl } amino) ) benzonitrile ¢© (hereinafter referred to as the compound (B tert-butyl 2-{7-[3-(4-cyanoanilino) propyl] -9-oxa-3, 7-diazabicyclo-[3,3. 1] non-1 \VoY yl } ethylcarbamate 7 -3 tert-butyl 2-{7-[(2S)-3-(4-cyanophenoxy)-2-hydroxypropyl]-9-oxa-3, 7 diazabicyclo A 1]non- 3-yl} ethylcarbamate A .3.3[ 0 (where this compound is hereinafter referred to as compound (C 1# -formulation according to claim 1 or ? or * or ; where The gelling polymer neutral Y is polyethylene oxide sli “, cult ¥ polyethylene glycol, or a mixture of two or more polyethylene oxides .polyethylene oxides ¢ 1 + 1 -formulation according to Claims 1 or JY ? or; wherein the ¥ neutral gel polymer is <hydroxy propyl methyl cellulose or a mixture of two or more different hydroxy propyl methyl celluloses ¢. 1 7 -- formatting according to claims 1 or 7 or © or ; Wherein, the ¥ neutral gel polymer is a mixture of hydroxy propyl methyl celluloses Vv and polyethylene oxide. where the ratio of the neutral gel polymer to iota-carrageenan is in the range from 0:01 to 860 Yer 401 - a formula according to what is mentioned in any of the previous claims, where ¥ the pharmaceutically active base ingredient is metoprolol. 1 wherein this process involves mixing iota-carrageenan ¢, one or more ¥ neutralizing gel polymers, and a basic pharmaceutical active ingredient. 11 1- Using the formula according to what is stated in Claim 1 in the manufacture of a remedy. 1 1 - Using the formula according to what is mentioned in Claim 1 in the manufacture of Y treatment for cardiovascular disorder. Dla