Fast-acting insulin composition comprising a substituted anionic compound and a polyanionic compound

Abstract

A composition, in the form of an aqueous solution, including insulin in hexameric form, at least one substituted anionic compound of non-saccharide structure and at least one polyanionic compound other than the substituted anionic compound.

Claims

1. A composition, in the form of an aqueous solution, comprising insulin in hexameric form, at least one substituted anionic compound of non-saccharide structure and at least one polyanionic compound other than said substituted anionic compound, wherein the substituted anionic compound corresponds to formula I below: ##STR00049## wherein R represents a saturated or unsaturated, linear, branched or cyclic hydrocarbon-based radical comprising from 1 to 12 carbon atoms, optionally comprising at least one function chosen from ether, alcohol and carboxylic acid functions, AA is a radical resulting from an aromatic amino acid comprising a phenyl group or an indole group, which is substituted or not substituted, or an aromatic amino acid derivative comprising a phenyl group or an indole group, which is substituted or not substituted, said radical AA bearing at least one free acid function, E represents an at least divalent radical, comprising from 2 to 6 carbon atoms, F, F′ and F″ represent, independently of each other, a function chosen from amide, carbamate and urea functions, F and F″ being functions resulting from a reaction involving the amine of the aromatic amino acid, the precursor of the radical AA, F′ being a function involving a reactive function of the precursor of R and a reactive function of the precursor of E, p being an integer between 1 and 3, m is an integer between 0 and 6; n is an integer between 0 and 6; m+n is an integer between 1 and 6; said compound comprising at least two carboxylic acid functions in the form of a salt of an alkali metal chosen from Na.sup.+ and K.sup.+.

2. The composition as claimed in claim 1, wherein the substituted anionic compound is chosen from the compounds of formula I wherein the radical —R— is chosen from radicals comprising 1 to 12 carbon atoms, of formula IV: ##STR00050## wherein q and r are integers between 0 and 12, r is an integer between 0 and 3 and 1≦q+r*t≦12, a is equal to 0 or 1, the groups R.sub.q1, R.sub.q2, R.sub.r1 and R.sub.r2 are, independently of each other, chosen from H, —OH and —COOH, if a=0, then t=0 and when q≧1 and/or t≧1, then the radicals R.sub.q1 and R.sub.q2 and the radicals R.sub.r1 and R.sub.r2 are identical or different from one carbon to another.

3. The composition as claimed in claim 1, wherein the substituted anionic compound is chosen from the compounds of formula I wherein the radical AA is resulting from an aromatic amino acid comprising a phenyl or an indole, which is substituted or not substituted, or from an aromatic amino acid derivative comprising a phenyl or an indole, which is substituted or not substituted, most particularly, the radical AA is resulting from an aromatic amino acid comprising a phenyl or an indole, which is substituted or not substituted.

4. The composition as claimed in claim 1, wherein the substituted anionic compound is chosen from the compounds of formula I wherein the radical E is resulting from a linear or branched alkyl compound comprising at least two functions chosen from the group consisting of OH, —COOH and —NH.sub.2.

5. The composition as claimed in claim 1, wherein the substituted anionic compound is chosen from the compounds of formula I wherein n=0 and corresponds to formula II: ##STR00051## wherein: AA, F and R have the definitions given above, 1≦m≦6.

6. The composition as claimed in claim 1, wherein the substituted anionic compound is chosen from the compounds of formula I wherein m=0 and corresponds to formula III: ##STR00052## wherein: AA, E, F′, F″, p and R have the definitions given above, 1≦n≦6.

7. The composition as claimed in claim 1, wherein the mole ratios of substituted anionic compound/insulin are between 0.6 and 120.

8. The composition as claimed in claim 1, wherein the mass ratios of substituted anionic compound/insulin are between 0.5 and 10.

9. The composition as claimed in claim 1, wherein the concentration of substituted anionic compound is between 1.8 and 36 mg/mL.

10. The composition as claimed in claim 1, wherein the insulin is human insulin chosen from recombinant human insulins.

11. The composition as claimed in claim 1, wherein insulin is an insulin analogue chosen from the group consisting of lispro insulin (Humalog®), aspart insulin (Novolog®, Novorapid®) and glulisine insulin (Apidra®).

12. A composition comprising a composition as claimed in claim 1, wherein the insulin concentration is between 240 and 3000 μM (40 to 500 IU/mL).

13. The composition as claimed in claim 1, wherein the polyanionic compound is chosen from the group consisting of polycarboxylic acids and the Na.sup.+, K.sup.+, Ca.sup.2+ or Mg.sup.2+ salts thereof.

14. The composition as claimed in claim 1, wherein the polyanionic compound is an anionic molecule chosen from the group consisting of citric acid, aspartic acid, glutamic acid, malic acid, tartaric acid, succinic acid, adipic acid, oxalic acid, phosphate, polyphosphoric acids and the Na.sup.+, K.sup.+, Ca.sup.2+ or Mg.sup.2+ salts thereof.

15. The composition as claimed in claim 1, wherein the concentration of polyanionic compound is between 2 and 150 mM.

16. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical AA is resulting from phenylalanine and the radical R is resulting from tartaric acid, succinic acid or an amino acid chosen from aspartic acid and glutamic acid, the radical R is especially resulting from tartaric acid or succinic acid and in particular n=0 and m=1.

17. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical AA is resulting from phenylalanine and the radical R is resulting from succinic acid, tartaric acid or an amino acid chosen from aspartic acid and glutamic acid, n=0 and m=1.

18. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical AA is resulting from phenylalanine, n=0, m=1, the radical R is resulting from tartaric acid or succinic acid, in particular from tartaric acid, and bears an acid function.

19. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical AA is resulting from phenylalanine, m=0, n=1 or 2, especially n=1, p=3 and E is resulting from TRIS.

20. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical AA is resulting from phenylalanine, m=0, n=1 or 2, especially n=1, p=2 and E is resulting from aspartic acid or glutamic acid.

21. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical AA is resulting from phenylalanine, m=0, n=1, 2 or 3, especially n=3, p=2 and E is resulting from aspartic acid or glutamic acid.

22. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical R is resulting from succinic acid and comprises a carboxylic acid, m=0, n=1, p=3, E is resulting from TRIS, F′ is an amide function, the radical AA is resulting from phenylalanine, and F″ is a carbamate function.

23. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical R is resulting from succinic acid and comprises a carboxylic acid, m=0, n=1, p=3, E is resulting from TRIS, F′ is an amide function, the radical AA is resulting from phenylalanine, and F″ is a urea function.

24. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical R is resulting from tartaric acid and comprises a carboxylic acid, m=1, n=0, F is an amide function and the radical AA is resulting from phenylalanine.

25. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical R is resulting from succinic acid and comprises a carboxylic acid, m=1, n=0, F is an amide function and the radical AA is resulting from phenylalanine.

26. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical R is resulting from tartaric acid and comprises a carboxylic acid, m=0, n=1, p=3, E is resulting from TRIS, F′ is an amide function, the radical AA is resulting from phenylalanine, and F″ is a carbamate function.

27. The composition as claimed in claim 1, wherein the substituted anionic compound corresponds to formula I wherein the radical R is resulting from tartaric acid and comprises a carboxylic acid, m=0, n=1, p=2, E is resulting from aspartic acid, F′ is an amide function, the radical AA is resulting from phenylalanine, and F″ is an amide function.

Description

BRIEF DESCRIPTION OF DRAWINGS

(1) FIG. 1 describes the pharmacodynamic results obtained with the compositions described in examples B2 and B12.

(2) FIG. 2 describes the pharmacokinetic results obtained with the compositions described in examples B2 and B12.

(3) FIG. 3 describes the pharmacodynamic results obtained with the compositions described in examples B2 and B7.

(4) FIG. 4 describes the pharmacokinetic results obtained with the compositions described in examples B2 and B7.

(5) FIG. 5 describes on the y-axis the CD signal at 276 nm (deg.Math.cm.sup.2.Math.dmol.sup.−1) and, on the x-axis:

(6) A: rhINS (human insulin) (100 IU/ml)

(7) B: rhINS/EDTA

(8) C: rhINS/Citrate

(9) D: rhINS/EDTA/Citrate

(10) E: rhINS/Compound A1 (100 IU/ml/7.3 mg/mL)

(11) FIG. 6 describes on the y-axis the CD signal at 276 nm (deg.Math.cm.sup.2.Math.dmol.sup.−1) and, on the x-axis:

(12) F: rhINS (100 IU/ml)

(13) G: rhINS/Compound A2 (100 IU/ml/7.3 mg/mL)

(14) H: rhINS/Compound A3 (100 IU/ml/7.3 mg/mL)

(15) FIG. 7 describes the pharmacodynamic results obtained with the compositions described in examples B2 and B39.

(16) FIG. 8 describes the pharmacokinetic results obtained with the compositions described in examples B2 and B39.

(17) FIG. 9 describes the pharmacodynamic results obtained with the compositions described in examples B2 and B47.

(18) FIG. 10 FIG. 10 describes the pharmacokinetic results obtained with the compositions described in examples B2 and B47.

(19) FIG. 11 describes the pharmacodynamic results obtained with the compositions described in examples B2 and B55.

(20) FIG. 12 describes the pharmacokinetic results obtained with the compositions described in examples B2 and B55.

(21) FIG. 13 describes on the y-axis the CD signal at 240 nm (deg.Math.cm.sup.2.Math.dmol.sup.−1) and, on the x-axis:

(22) A: lispro insulin 100 IU/mL

(23) B: lispro insulin+7.3 mg/mL of substituted anionic compound A1+citrate at 9.3 mM

(24) C: lispro insulin+7.3 mg/mL of substituted anionic compound A2+citrate at 9.3 mM

(25) D: lispro insulin+7.3 mg/mL of substituted anionic compound A5+citrate at 9.3 mM

(26) E: lispro insulin+7.3 mg/mL of substituted anionic compound A6+citrate at 9.3 mM

(27) F: lispro insulin+7.3 mg/mL of substituted anionic compound A7+citrate at 9.3 mM

(28) G: lispro insulin+EDTA 300 μM

(29) H: lispro insulin+citrate at 9.3 mM

(30) FIG. 14 describes the pharmacodynamic results obtained with the compositions described in examples B2 and B7.

(31) FIG. 15 describes the pharmacokinetic results obtained with the compositions described in examples B2 and B7.

EXAMPLES

(32) TABLE-US-00001 TABLE 1 below presents, in a nonlimiting manner, examples of compounds according to the invention. Substituted anionic compounds Formulations A1 A2 A3 A4 A5 A6 A7
Part A: Synthesis of the Substituted Anionic Compounds

Example A1

Substituted Anionic Compound A1

(33) The substituted anionic compound A1 or N-(3-carboxy-1-oxopropyl-)-L-phenylalanine is obtained according to a modification of the process described in patent application WO 96/33699 (Milstein, S.) from L-phenylalanine ethyl ester and O,O′-diacetyltartaric anhydride.

(34) Yield: 6.7 g (26%)

(35) .sup.1H NMR (DMSO-d.sub.6, ppm): 3.10 (2H); 4.20 (1H); 4.30 (1H); 4.50 (1H); 7.25 (5H); 7.70 (1H).

Example A2

Substituted Anionic Compound A2

Molecule 1

L-Phenylalanine Ethyl Ester Isocyanate

(36) To a solution of the hydrochloride salt of L-phenylalanine ethyl ester (23 g, 100 mmol) in a mixture of dichloromethane (400 mL) and saturated aqueous sodium hydrogen carbonate solution (400 mL) at 0° C. is added one portion of triphosgene (9.8 g, 33 mmol). After stirring for 1 hour at 0° C., the organic phase is separated out and the aqueous phase is extracted 3 times with dichloromethane. The combined organic phases are dried over Na.sub.2SO.sub.4, filtered and concentrated to give a colorless oil which gradually crystallizes.

(37) Yield: 22 g (quantitative)

(38) .sup.1H NMR (CDCl.sub.3, ppm): 1.24 (3H); 2.98 (1H); 3.11 (1H); 4.17-4.23 (3H); 7.14-7.28 (5H).

Molecule 2

Product Obtained by Reaction Between Tris(Hydroxymethyl)Aminomethane and Monomethyl Succinate

(39) Via a process similar to that described in J. Org. Chem. 2011, 76, 2084 by reaction between tris(hydroxymethyl)aminomethane (2.66 g, 22 mmol) and monomethyl succinate (2.64 g, 20 mmol), a white solid is obtained.

(40) Yield: 3.77 g (80%)

(41) .sup.1H NMR (CDCl.sub.3, ppm): 2.52 (2H); 2.52 (2H); 3.64-3.70 (9H); 6.69 (1H).

Molecule 3

Product Obtained by Reaction Between Molecule 2 and Molecule 1

(42) 1,4-Diazobicyclo[2.2.2]octane (DABCO) (136 mg, 1.21 mmol) is added, under nitrogen, to a solution of molecule 2 (1.68 g, 7.14 mmol) and molecule 1 (5.63 g, 25.7 mmol) in toluene (70 mL), and the mixture is heated at 90° C. for 2.5 days. After concentrating the reaction medium under vacuum and coevaporation of the toluene with ethanol, the residue obtained is taken up in dichloromethane and washed with 1N HCl. The aqueous phase is extracted twice with dichloromethane. The organic phases are combined, dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum. Purification by flash chromatography (cyclohexane/ethyl acetate) leads to the desired product in the form of a colorless oil.

(43) Yield: 5.15 g (80%)

(44) .sup.1H NMR (CDCl.sub.3, ppm): 1.24 (9H); 2.39 (2H); 2.59 (2H); 2.90-3.15 (6H); 3.63 (3H); 4.10-4.30 (9H); 4.45-4.60 (6H); 6.07 (3H); 7.10-7.35 (15H).

(45) Substituted Anionic Compound A2

(46) A suspension of molecule 3 (10.2 g, 11.43 mmol) in an ethanol/tetrahydrofuran (THF)/water mixture (50 mL/32 mL/32 mL) is treated under nitrogen with 2N NaOH (23.4 mL) and then stirred at room temperature overnight. The volatile solvents are evaporated off under vacuum and 25 mL of 2N HCl are then gradually added to the medium. The medium is then extracted with ethyl acetate and the organic phase is dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a foam. After triturating in pentane and ethyl ether, filtering and drying under vacuum, a white issue solid corresponding to the anionic molecule A2 in acid form is obtained. The solid is dissolved in a water/1-butanol mixture (50/50 vol/vol) and lyophilized.

(47) Yield: 5.9 g (86%)

(48) .sup.1H NMR (DMSO-d.sub.6, ppm): 2.10-2.20 (4H); 2.80-3.10 (6H); 4.00-4.20 (9H); 7.10-7.30 (15H); 7.50-7.60 (4H); 12.20-12.80 (4H).

(49) LC/MS (ESI): 795.3; (calculated ([M+H].sup.+): 795.3).

(50) The substituted anionic compound A2 in acid form is dissolved in water and the solution is neutralized by gradual addition of 10N sodium hydroxide to give an aqueous solution of substituted anionic compound A2, which is then lyophilized.

(51) .sup.1H NMR ((D.sub.2O, ppm): 2.25-2.40 (4H); 2.75-2.90 (3H); 3.10-3.30 (3H); 3.75-4.30 (9H); 7.10-7.40 (15H).

Example A3

Substituted Anionic Compound A3

Molecule 4

Product Obtained by Reaction Between the Hydrochloride Salt of L-Phenylalanine Ethyl Ester and N-(Tert-Butoxycarbonyl)-L-Aspartic Acid

(52) A suspension of N-(tert-butoxycarbonyl)-L-aspartic acid (2.33 g, 10 mmol) in 120 mL of dichloromethane at 0° C. is treated, under nitrogen, with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) (5.75 g, 30 mmol). After stirring for 35 minutes, the hydrochloride salt of L-phenylalanine ethyl ester (6.89 g, 30 mmol), 1-hydroxybenzotriazole (HOBt) (4.05 g, 30 mmol) and diisopropylethylamine (DIPEA) (3.7 mL) are added and the mixture is stirred at room temperature overnight. The medium is neutralized by adding 150 mL of saturated aqueous NaHCO.sub.3 solution and the medium is diluted with 150 mL of dichloromethane. The aqueous phase is extracted with dichloromethane. The organic phases are combined, washed with water, with saturated aqueous NaHCO.sub.3 solution and with saturated aqueous NH.sub.4Cl solution and then dried and concentrated under vacuum.

(53) Yield: 8.02 g (quantitative)

(54) .sup.1H NMR (CDCl.sub.3, ppm): 1.15-1.30 (6H); 1.42 (9H); 2.50-2.57 (1H); 2.83-2.89 (1H); 3.07-3.10 (4H); 4.05-4.22 (4H); 4.43 (1H); 4.73-4.83 (2H); 5.99-6.30 (1H); 6.27-6.29 (1H); 7.15-7.30 (10H).

Molecule 5

Hydrochloride Salt of the Product Obtained by Deprotection of the Boc Group of Molecule 4

(55) A solution of molecule 4 (8 g, 10 mmol) in dichloromethane at 0° C. is treated, under nitrogen, with 4N HCl solution in dioxane (26 mL). After 5 hours at 0° C., the medium is concentrated under vacuum and coevaporated several times with dichloromethane to give a viscous foam.

(56) Yield: 5.2 g (quantitative)

(57) .sup.1H NMR (DMSO-d.sub.6, ppm): 1.05-1.15 (6H); 2.49-2.68 (1H); 2.79-2.86 (1H); 2.95-3.05 (4H); 3.99-4.07 (5H); 4.40-4.50 (2H); 7.20-7.36 (10H); 8.16 (3H); 8.89 (1H); 8.99 (1H).

Molecule 6

Product Obtained by Reaction Between Succinic Acid and Molecule 5

(58) A suspension of succinic acid (0.33 g, 2.77 mmol) in dichloromethane (30 mL) at 0° C. is treated, under nitrogen, with EDCI (1.59 g, 8.3 mmol). After stirring for 35 minutes, a solution of molecule 5 (5.2 g, 10 mmol) in dichloromethane (10 mL), HOBt (1.12 g, 8.3 mmol) and DIPEA (1 mL) are added and the mixture is stirred at room temperature for 48 hours. The medium is diluted with dichloromethane and filtered through a sinter. The solid is washed with dichloromethane and then dried under vacuum to give a white solid.

(59) Yield: 2.9 g (quantitative)

(60) .sup.1H NMR (DMSO-d.sub.5, ppm): 0.95-1.15 (12H); 2.20-2.40 (8H); 2.80-3.10 (8H); 3.90-4.10 (8H); 4.30-4.50 (4H); 4.50-4.65 (4H); 7.10-7.40 (20H); 8.03 (2H); 8.13 (2H); 8.32 (2H).

(61) Substituted Anionic Compound A3

(62) A suspension of molecule 6 (2.9 g, 2.77 mmol) in a THF/ethanol/water mixture (50/50/25 mL) is treated, under nitrogen, with aqueous 2N NaOH solution (5.68 mL). After stirring at room temperature overnight, the reaction medium is filtered through a sinter and the filtrate is concentrated under vacuum. The aqueous phase obtained is acidified with 2N HCl and the precipitate formed is filtered off and then transferred into a round-bottomed flask with methanol. The solution is concentrated under vacuum and dried under vacuum to give a white solid of the substituted anionic compound A3 in acid form.

(63) Yield: 1.98 g (76%)

(64) .sup.1H NMR (DMSO-d.sub.6, ppm): 2.20-2.60 (8H); 2.80-3.10 (8H); 3.80-4.10 (8H); 4.30-4.45 (4H); 4.50-4.60 (2H); 7.10-7.35 (20H); 7.91 (2H); 8.06 (2H); 8.25 (2H).

(65) LC/MS (ESI): 937.5; (calculated ([M+H].sup.+): 937.9).

(66) The substituted anionic compound A3 in acid form is dissolved in water and the solution is neutralized by gradual addition of 10N sodium hydroxide to give an aqueous solution of anionic molecule A3, which is then lyophilized.

(67) .sup.1H NMR ((D.sub.2O, ppm): 2.20-2.25 (4H); 2.45-2.55 (2H); 2.60-2.75 (2H); 2.75-3.00 (4H); 3.15-3.25 (4H); 4.30-4.45 (4H); 4.55-4.80 (2H); 7.10-7.35 (20H).

Example A4

Substituted Anionic Compound A4

Molecule 7

Product Obtained by Reaction Between Diethyl Succinate and Tris(Hydroxymethyl)Aminomethane

(68) A suspension of diethyl succinate (8.72 g, 50 mmol) and of tris(hydroxymethyl)aminomethane (12.72 g, 105 mmol) in an ethanol/water mixture (10/1 vol/vol) is heated, under nitrogen, at 100° C. for 7 hours and then at 60° C. for 2 days. The white precipitate formed is filtered off on a sinter, rinsed with ethanol and then dried under vacuum. The solid is taken up in an ethanol/water mixture (9/1 by vol) and then filtered off, rinsed with an ethanol/water mixture (9/1 by vol) and dried under vacuum.

(69) Yield: 7.1 g (44%)

(70) .sup.1H NMR (DMSO-d.sub.6, ppm): 2.54 (4H); 3.51 (12H); 4.65 (6H); 7.16 (2H).

Molecule 8

Product Obtained by Reaction Between Molecule 7 and Molecule 1

(71) A suspension of molecule 7 (9.9 g, 30.5 mmol) and of molecule 1 (47 g, 213 mmol) in toluene is treated, under nitrogen, with 1,4-diazobicyclo[2.2.7]octane (DABCO) (1.37 g, 12.2 mmol), and the mixture is heated at 110° C. for 6 hours and then at 90° C. overnight. The medium is concentrated under vacuum and then taken up in 750 mL of ethyl acetate and 250 mL of 1N HCl. The organic phase is washed with saturated NaCl solution, dried and concentrated under vacuum. The residue is purified by flash chromatography (cyclohexane/ethyl acetate).

(72) Yield: 21.1 g (42%)

(73) .sup.1H NMR (CDCl.sub.3, ppm): 1.22 (18H); 2.35 (4H); 2.90-3.15 (12H); 4.15-4.30 (18H); 4.40-4.65 (12H); 6.07 (6H); 7.10-7.35 (30H).

(74) Anionic Molecule A4

(75) A solution of molecule 8 (21.1 g, 12.8 mmol) in an ethanol/THF/water mixture (48/48/24 mL) is treated, under nitrogen, with aqueous 2N NaOH solution (42.2 mL). After stirring at room temperature for 24 hours, the reaction medium is concentrated under vacuum and the aqueous phase is acidified with 2N HCl. The precipitate is filtered off, rinsed with water and then taken up in water. The suspension is frozen and then lyophilized to give a white solid of anionic molecule A4 in acid form.

(76) Yield: 18 g (95%)

(77) .sup.1H NMR (DMSO-d.sub.6, ppm): 2.19 (4H); 2.75-3.10 (12H); 3.80-4.30 (18H); 7.10-7.30 (30H); 7.51 (8H).

(78) LC/MS (ESI): 734.8; (calculated ([M/2]): 735.7).

(79) The substituted anionic compound A4 in acid form is dissolved in water and the solution is neutralized by gradual addition of 10N sodium hydroxide to give an aqueous solution of anionic molecule A4, which is then lyophilized.

(80) .sup.1H NMR ((D.sub.2O, ppm): 2.20 (4H); 2.75-2.95 (6H); 3.10-3.30 (6H); 3.80-4.35 (18H); 7.10-7.30 (30H).

Example A5

Substituted Anionic Compound A5

(81) Triethylamine (30.6 g, 0.3 mol) is added to a solution of L-phenylalanine (25 g, 0.15 mol) in THF (200 mL) at room temperature, followed by addition of a solution of succinic anhydride (15.1 g, 0.15 mol) in THF (189 mL). The reaction medium is brought to the boiling point and heated for 16 hours. After cooling to room temperature, the medium is concentrated under vacuum to three-quarters of its volume and then acidified to pH 1.5 by adding 12N HCl. The aqueous phase is extracted with ethyl acetate. The organic phase is washed with 0.01N HCl and concentrated under vacuum to give a viscous yellow oil. This oil is taken up in water and the solution is stirred at pH 12 for 3 hours following the addition of 10N sodium hydroxide. The solution is then neutralized and then acidified by adding 12N HCl to pH 1.5. The solution is extracted with ethyl acetate, concentrated under vacuum and dried under vacuum to give a white solid of the substituted anionic compound A5 in acid form.

(82) Yield: 26.9 g (67%)

(83) .sup.1H NMR (DMSO-d.sub.5, ppm): 2.33 (4H); 2.75-3.15 (2H); 4.42 (1H); 7.21 (5H); 8.18 (1H); 12.38 (2H).

(84) LC/MS (ESI): 266.2; (calculated ([M+H].sup.+): 266.3).

(85) The substituted anionic compound A5 in acid form is dissolved in water and the solution is neutralized by gradual addition of 10N sodium hydroxide to give an aqueous solution of substituted anionic compound A5 or N-(β-carboxypropionyl)-L-phenylalanine, which is then lyophilized.

(86) .sup.1H NMR ((D.sub.2O, ppm): 2.25-2.45 (4H); 2.65-2.92 (1H); 3.08-3.19 (1H); 4.35-4.45 (1H); 7.05-7.39 (5H)

Example A6

Substituted Anionic Compound A6

Molecule 9

Carbamic acid N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]-1,1-dimethylethyl ester

(87) Molecule 9 is obtained according to the process described in patent WO 2008/30119 starting with 20 g of tris(hydroxymethyl)aminomethane.

(88) Yield: 33.2 g (88%)

(89) .sup.1H NMR (DMSO-d.sub.5, ppm): 1.37 (9H); 3.50 (6H); 4.48 (3H); 5.76 (1H).

Molecule 10

Product Obtained by Reaction Between Molecule 1 and Molecule 9

(90) 1,4-Diazobicyclo[2.2.7]octane (DABCO) (795 μL, 7.2 mmol) is added to a solution of molecule 9 (4 g, 18.08 mmol) and of molecule 1 (11.87 g, 57.9 mmol) in toluene (260 mL), and the mixture is heated at 110° C. for 3 hours and then at 90° C. overnight. After concentrating under vacuum, the medium is taken up in ethyl acetate and then washed with 1N HCl. The organic phase is washed with saturated NaCl solution, dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum. The residue is purified by flash chromatography (cyclohexane/ethyl acetate).

(91) Yield: 7.5 g (50%)

(92) .sup.1H NMR (CDCl.sub.3, ppm): 1.41 (9H); 2.85-3.20 (6H); 3.73 (9H); 4.25 (3H); 4.39 (3H); 4.57 (3H); 5.72 (3H); 7.10-7.35 (15H).

(93) LC/MS (ESI): 837.7; (calculated ([M+H].sup.+): 837.8).

Molecule 11

Hydrochloride Salt of the Product Obtained by Deprotection of the Boc Group of Molecule 10

(94) A solution of molecule 10 (7.5 g, 9 mmol) in dichloromethane at 0° C. is treated, under argon, with 4N HCl solution in dioxane (22.4 mL). The medium is stirred from 0° C. to room temperature for 3 hours and is then concentrated under vacuum to give a white gum, which is dried under vacuum at room temperature for 15 hours.

(95) Yield: 6.84 g (99%) .sup.1H NMR (CDCl.sub.3, ppm): 3.02-3.20 (6H); 3.73 (9H); 4.19 (3H); 4.42 (3H); 4.55 (3H); 5.72 (3H); 7.15-7.35 (15H); 8.93 (3H).

(96) LC/MS (ESI): 737.4; (calculated ([M-Cl.sup.−]): 737.7).

Molecule 12

Product Obtained by Reaction Between Molecule 11 and O,O′-Diacetyltartaric Anhydride

(97) Molecule 11 (5.7 g, 7.4 mmol) is dissolved in dichloromethane (50 mL) and saturated NaHCO.sub.3 solution (30 mL) is added. The aqueous phase is extracted with dichloromethane and the organic phase is dried over Na.sub.2SO.sub.4, filtered and concentrated to a volume of about 20 mL. This solution is poured onto a solution of 0,0′-diacetyltartaric anhydride (1.6 g, 7.4 mmol) in THF (35 mL) at 0° C. After stirring for 4 hours, the medium is concentrated under vacuum to give a white foam. The product is purified by flash chromatography (dichloromethane/methanol) to give a white foam.

(98) Yield: 5.5 g (78%)

(99) .sup.1H NMR (CDCl.sub.3, ppm): 2.05 (3H); 2.15 (3H); 2.93 (3H); 3.05 (3H); 3.73 (9H); 4.19 (3H); 4.50 (6H); 5.60 (1H); 5.69 (1H); 6.30 (3H); 7.15-7.35 (15H); 7.92 (1H).

(100) LC/MS (ESI): 953.6; (calculated ([M+H].sup.+): 953.9).

Example A6

Substituted Anionic Compound A6

(101) To a solution of molecules 12 (5.14 g, 5.39 mmol) in a methanol/water/THF mixture (18 mL/18 mL/18 mL) at 0° C. is added aqueous 2N sodium hydroxide solution (13.5 mL) and the mixture is stirred at 0° C. for 2 hours. After removal of the methanol and THF by evaporation under vacuum and washing of the solution with ethyl acetate, the solution is cooled to 0° C. and acidified to pH 1 by adding aqueous 10% HCl solution. The precipitated product is extracted with ethyl acetate 3×100 mL). The combined organic phases are dried over Na2SO4, filtered and concentrated under vacuum to give a white solid. The residue is taken up twice in a methanol/toluene mixture and then evaporated to dryness under vacuum. The solid is suspended in water and the mixture is lyophilized.

(102) Yield: 3.8 g (85%)

(103) LC/MS (ESI): 827.4; (calculated ([M+H].sup.+): 827.7).

(104) The substituted anionic compound A6 (3.8 g) in acid form is suspended in water and 1.0025 M sodium hydroxide is gradually added to pH 7.18. After lyophilization, the desired product is obtained in the form of a white solid.

(105) Yield: 4.09 g (83%)

(106) .sup.1H NMR (D.sub.2O, 80° C., ppm): 3.45 (3H); 3.75 (3H); 4.55-4.75 (6H); 4.75-4.85 (4H); 5.00 (1H); 7.75-8.00 (15H).

(107) LC/MS (ESI): 827.5; (calculated ([M+3H+−3Na+]): 827.7).

Example A7

Substituted Anionic Compound A7

Molecule 13

Product Obtained by Reaction Between Molecule 5 and O,O′-Diacetyltartaric Anhydride

(108) An aqueous solution (60 mL) of molecule 5 (4.99 g, 9.6 mmol) is poured into saturated NaHCO.sub.3 solution (60 mL) and the mixture is extracted with dichloromethane (3×30 mL). The organic phase is dried over Na.sub.2SO.sub.4, filtered and concentrated to two-thirds of its volume under vacuum. It is added to a solution of O,O′-diacetyltartaric anhydride (2.07 g, 9.6 mmol) in THF at 0° C. and the mixture is stirred at 0° C. for 30 minutes. The mixture is concentrated under vacuum to obtain a white foam.

(109) Yield: 6.45 g (96%)

(110) .sup.1H NMR (DMSO-d.sub.6, ppm): 1.01-1.10 (6H); 2.00 (3H); 2.10 (3H); 2.43 (4H); 2.80-2.95 (4H); 3.60 (1H); 3.95-4.05 (4H); 4.39 (2H); 4.60 (1H); 5.50 (2H); 7.19-7.30 (10H); 8.25-8.40 (3H); 13.50 (1H).

(111) LC/MS (ESI): 700.3; (calculated ([M+H].sup.+): 700.7).

(112) Substituted Anionic Compound A7

(113) To a solution of molecule 13 (5.39 g, 7.70 mmol) in a methanol/water mixture (38 mL/38 mL) at 0° C. is added aqueous 2N sodium hydroxide solution (15.4 mL) and the mixture is stirred at 0° C. for 1 hour. The methanol is removed under reduced pressure and the volume of water is reduced to about one third. The medium is acidified at 0° C. by slow addition of aqueous 10% HCl solution and is then extracted with ethyl acetate (3×50 mL). The organic phases are dried over Na.sub.2SO.sub.4 and evaporated under reduced pressure to give a white solid of the substituted anionic compound A7 in acid form, which is treated azeotropically 3 times with 50 mL of water to remove the traces of ethyl acetate and of acetic acid.

(114) Yield: 3.24 g (75%)

(115) .sup.1H NMR (DMSO-d.sub.6, ppm): 2.55 (1H); 2.65 (1H); 2.80-3.05 (4H); 3.35 (1H); 4.25-4.45 (4H); 4.58 (1H); 5.93 (1H); 7.19-7.30 (10H); 7.81 (1H); 8.10 (1H); 8.28 (1H); 12.65 (3H).

(116) LC/MS (ESI): 560.1; (calculated ([M+H].sup.+): 560.5).

(117) The substituted anionic compound A7 (3.24 g) in acid form is suspended in 50 mL of water and 0.521 M sodium hydroxide is added dropwise to pH 7.08. After lyophilization, the desired product is obtained in the form of a white solid.

(118) Yield: 3.53 g (97%)

(119) .sup.1H NMR (MeOD-d.sub.4, ppm): 2.61 (1H); 2.70 (1H); 2.95 (2H); 3.15 (2H); 4.29 (1H); 4.42 (3H); 4.64 (1H); 7.10-7.30 (10H).

(120) LC/MS (ESI): 560.2; (calculated ([M+3H+−3Na+]): 559.4).

(121) Part A′: Synthesis of a Polyanionic Compound

(122) Polyanionic Compound 1: Sodium Maltotriosemethylcarboxylate

(123) 0.6 g (16 mmol) of sodium borohydride is added to 8 g (143 mmol of hydroxyl functions) of maltotriose (CarboSynth) dissolved in water at 65° C. After stirring for 30 minutes, 28 g (238 mmol) of sodium chloroacetate are added. 24 mL of 10 N NaOH (240 mmol) are then added dropwise to this solution, and the mixture is then heated at 65° C. for 90 minutes. 16.6 g (143 mmol) of sodium chloride acetate are then added to the reaction medium, along with dropwise addition of 14 mL of 10 N NaOH (140 mmol). After heating for 1 hour, the mixture is diluted with water, neutralized with acetic acid and then purified by ultrafiltration on a 1 kDa PES membrane against water. The concentration of compound in the final solution is determined from the dry extract, and an acid/base titration in a 50/50 (V/V) water/acetone mixture is then performed to determine the degree of substitution with sodium methylcarboxylate.

(124) According to the dry extract: [polyanionic compound 1]=32.9 mg/g

(125) According to the acid/base titration: the degree of substitution with sodium methylcarboxylates per saccharide unit is 1.65.

(126) The polyanionic compounds are selected by measuring their dissociation constant with respect to calcium ions and with respect to their capacity for not destabilizing the hexameric form of insulin.

(127) As regards the dissociation constant with respect to calcium ions, it is determined as follows.

(128) Solutions containing 2.5 mM of CaCl.sub.2, 150 mM of NaCl and increasing concentrations (between 0 and 20 mM) of polyanionic compound are prepared. The potential of all these formulations is measured and the concentrations of free calcium ions in the formulations are determined. After linearization by the Scatchard method, the dissociation constants are established. These data make it possible to compare the affinity of the various polyanionic compounds for Ca.

(129) Part B: Preparation of the Solutions

(130) B1. Novolog® Rapid Insulin Analog Solution at 100 IU/mL

(131) This solution is a commercial solution of aspart insulin from Novo Nordisk sold under the name Novolog®. This product is a rapid insulin analog.

(132) B2. Humalog® Rapid Insulin Analog Solution at 100 IU/mL

(133) This solution is a commercial solution of lispro insulin from Eli Lilly sold under the name Humalog®. This product is a rapid insulin analog.

(134) B3. Humulin® R Human Insulin Solution at 100 IU/mL

(135) This solution is a commercial solution of human insulin from Eli Lilly sold under the name Humulin® R. This product is a human insulin composition.

(136) B4. Apidra® Rapid Insulin Analog Solution at 100 IU/mL

(137) This solution is a commercial solution of glulisine insulin from Sanofi sold under the name Apidra®. This product is a rapid insulin analog.

(138) B5. Preparation of a 1.188 M Sodium Citrate Solution

(139) The sodium citrate solution is obtained by dissolving 9.0811 g of sodium citrate (30.9 mmol) in 25 mL of water in a graduated flask. The pH is adjusted to 7.4 by adding 1 mL of 1 M HCl. The solution is filtered through a 0.22 μm membrane.

(140) B7. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(141) For a final volume of 100 mL of composition, with a [substituted anionic compound A1]/[lispro insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(142) TABLE-US-00002 Lyophilized compound (substituted anionic compound A1) 730 mg Commercial solution of Humalog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(143) The final pH is adjusted to 7.4±0.4.

(144) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(145) B8. Preparation of a Solution of Human Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(146) For a final volume of 100 mL of composition, with a [substituted anionic compound A1]/[human insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(147) TABLE-US-00003 Lyophilized compound (substituted anionic compound A1) 730 mg Commercial solution of Humulin ® R 100 mL Sodium citrate solution at 1.188M 783 μL

(148) The final pH is adjusted to 7.4±0.4.

(149) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(150) B9. Preparation of a Solution of Aspart Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(151) For a final volume of 100 mL of composition, with a [substituted anionic compound A1]/[aspart insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(152) TABLE-US-00004 Lyophilized compound (substituted anionic compound A1) 730 mg Commercial solution of Novolog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(153) The final pH is adjusted to 7.4±0.4.

(154) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(155) B10. Preparation of a Solution of Glulisine Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(156) For a final volume of 100 mL of composition, with a [substituted anionic compound A1]/[glulisine insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(157) TABLE-US-00005 Lyophilized compound (substituted anionic compound A1) 730 mg Commercial solution of Apidra ® 100 mL Sodium citrate solution at 1.188M 783 μL

(158) The final pH is adjusted to 7.4±0.4.

(159) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(160) B12. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(161) For a final volume of 100 mL of composition, with a [substituted anionic compound A2]/[lispro insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(162) TABLE-US-00006 Lyophilized compound (substituted anionic compound A2) 730 mg Commercial solution of Humalog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(163) The final pH is adjusted to 7.4±0.4.

(164) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(165) B13. Preparation of a Solution of Human Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(166) For a final volume of 100 mL of composition, with a [substituted anionic compound A2]/[human insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(167) TABLE-US-00007 Lyophilized compound (substituted anionic compound A2) 730 mg Commercial solution of Humulin ® R 100 mL Sodium citrate solution at 1.188M 783 μL

(168) The final pH is adjusted to 7.4±0.4.

(169) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(170) B14. Preparation of a Solution of Aspart Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(171) For a final volume of 100 mL of composition, with a [substituted anionic compound A2]/[aspart insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(172) TABLE-US-00008 Lyophilized compound (substituted anionic compound A2) 730 mg Commercial solution of Novolog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(173) The final pH is adjusted to 7.4±0.4.

(174) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(175) B15. Preparation of a Solution of Glulisine Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(176) For a final volume of 100 mL of composition, with a [substituted anionic compound A2]/[glulisine insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(177) TABLE-US-00009 Lyophilized compound (substituted anionic compound A2) 730 mg Commercial solution of Apidra ® 100 mL Sodium citrate solution at 1.188M 783 μL

(178) The final pH is adjusted to 7.4±0.4.

(179) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(180) B17. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(181) For a final volume of 100 mL of composition, with a [substituted anionic compound A3]/[lispro insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(182) TABLE-US-00010 Lyophilized compound (substituted anionic compound A3) 730 mg Commercial solution of Humalog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(183) The final pH is adjusted to 7.4±0.4.

(184) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(185) B18. Preparation of a Solution of Human Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(186) For a final volume of 100 mL of composition, with a [substituted anionic compound A3]/[human insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(187) TABLE-US-00011 Lyophilized compound (substituted anionic compound A3) 730 mg Commercial solution of Humulin ® R 100 mL Sodium citrate solution at 1.188M 783 μL

(188) The final pH is adjusted to 7.4±0.4.

(189) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(190) B19. Preparation of a Solution of Aspart Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(191) For a final volume of 100 mL of composition, with a [substituted anionic compound A3]/[aspart insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(192) TABLE-US-00012 Lyophilized compound (substituted anionic compound A3) 730 mg Commercial solution of Novolog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(193) The final pH is adjusted to 7.4±0.4.

(194) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(195) B20. Preparation of a Solution of Glulisine Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(196) For a final volume of 100 mL of composition, with a [substituted anionic compound A3]/[glulisine insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(197) TABLE-US-00013 Lyophilized compound (substituted anionic compound A3) 730 mg Commercial solution of Apidra ® 100 mL Sodium citrate solution at 1.188M 783 μL

(198) The final pH is adjusted to 7.4±0.4.

(199) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(200) B22. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(201) For a final volume of 100 mL of composition, with a [substituted anionic compound A4]/[lispro insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(202) TABLE-US-00014 Lyophilized compound (substituted anionic compound A4) 730 mg Commercial solution of Humalog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(203) The final pH is adjusted to 7.4±0.4.

(204) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(205) B23. Preparation of a Solution of Human Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(206) For a final volume of 100 mL of composition, with a [substituted anionic compound A4]/[human insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(207) TABLE-US-00015 Lyophilized compound (substituted anionic compound A4) 730 mg Commercial solution of Humalog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(208) The final pH is adjusted to 7.4±0.4.

(209) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(210) B24. Preparation of a Solution of Aspart Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(211) For a final volume of 100 mL of composition, with a [substituted anionic compound A4]/[aspart insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(212) TABLE-US-00016 Lyophilized compound (substituted anionic compound A4) 730 mg Commercial solution of Novolog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(213) The final pH is adjusted to 7.4±0.4.

(214) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(215) B25. Preparation of a Solution of Glulisine Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(216) For a final volume of 100 mL of composition, with a [substituted anionic compound A4]/[glulisine insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified in the table and in the following order

(217) TABLE-US-00017 Lyophilized compound (substituted anionic compound A4) 730 mg Commercial solution of Apidra ® 100 mL Sodium citrate solution at 1.188M 783 μL

(218) The final pH is adjusted to 7.4±0.4.

(219) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(220) B26. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A4 and of the Polyanionic Compound 1

(221) For a final volume of 100 mL of formulation, with a [compound A1]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/2.0/1, the various reagents are added in the amounts specified below and in the following order:

(222) TABLE-US-00018 Lyophilized compound A1 730 mg Lyophilized polyanionic compound 1 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(223) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(224) The final pH is adjusted to 7.4±0.4.

(225) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(226) B27. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A1 and of the Polyanionic Compound 1

(227) For a final volume of 100 mL of formulation, with a [compound A1]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are added in the amounts specified below:

(228) TABLE-US-00019 Compound A1 in lyophilized form 730 mg Polyanionic compound 1 in lyophilized form 1460 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(229) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(230) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(231) B28. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A4 and of the Polyanionic Compound 1

(232) For a final volume of 100 mL of formulation, with a [compound A1]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/5.5/1, the various reagents are added in the amounts specified below and in the following order:

(233) TABLE-US-00020 Lyophilized compound A1 730 mg Lyophilized polyanionic compound 1 2000 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(234) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(235) The final pH is adjusted to 7.4±0.4.

(236) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(237) B29. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A2 and of the Polyanionic Compound 1

(238) For a final volume of 100 mL of formulation, with a [compound A2]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/2.0/1, the various reagents are added in the amounts specified below and in the following order:

(239) TABLE-US-00021 Lyophilized compound A2 730 mg Lyophilized polyanionic compound 1 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(240) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(241) The final pH is adjusted to 7.4±0.4.

(242) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(243) B30. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A2 and of the Polyanionic Compound 1

(244) For a final volume of 100 mL of formulation, with a [compound A2]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are added in the amounts specified below:

(245) TABLE-US-00022 Compound A2 in lyophilized form 730 mg Polyanionic compound 1 in lyophilized form 1460 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(246) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(247) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(248) B31. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A2 and of the Polyanionic Compound 1

(249) For a final volume of 100 mL of formulation, with a [compound A2]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/5.5/1, the various reagents are added in the amounts specified below and in the following order:

(250) TABLE-US-00023 Lyophilized compound A2 730 mg Lyophilized polyanionic compound 1 2000 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(251) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(252) The final pH is adjusted to 7.4±0.4.

(253) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(254) B32. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A3 and of the Polyanionic Compound 1

(255) For a final volume of 100 mL of formulation, with a [compound A3]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/2.0/1, the various reagents are added in the amounts specified below and in the following order:

(256) TABLE-US-00024 Lyophilized compound A3 730 mg Lyophilized polyanionic compound 1 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(257) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(258) The final pH is adjusted to 7.4±0.4.

(259) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(260) B33. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A3 and of the Polyanionic Compound 1

(261) For a final volume of 100 mL of formulation, with a [compound A3]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are added in the amounts specified below:

(262) TABLE-US-00025 Compound A3 in lyophilized form 730 mg Polyanionic compound 1 in lyophilized form 1460 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(263) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(264) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(265) B34. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A3 and of the Polyanionic Compound 1

(266) For a final volume of 100 mL of formulation, with a [compound A3]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/5.5/1, the various reagents are added in the amounts specified below and in the following order:

(267) TABLE-US-00026 Lyophilized compound A3 730 mg Lyophilized polyanionic compound 1 2000 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(268) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(269) The final pH is adjusted to 7.4±0.4.

(270) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(271) B35. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A4 and of the Polyanionic Compound 1

(272) For a final volume of 100 mL of formulation, with a [compound A4]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/2.0/1, the various reagents are added in the amounts specified below and in the following order:

(273) TABLE-US-00027 Lyophilized compound A4 730 mg Lyophilized polyanionic compound 1 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(274) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(275) The final pH is adjusted to 7.4±0.4.

(276) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(277) B36. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A4 and of the Polyanionic Compound 1

(278) For a final volume of 100 mL of formulation, with a [compound A4]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are added in the amounts specified below:

(279) TABLE-US-00028 Compound A4 in lyophilized form 730 mg Polyanionic compound 1 in lyophilized form 1460 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(280) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(281) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(282) B37. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A4 and of the Polyanionic Compound 1

(283) For a final volume of 100 mL of formulation, with a [compound A4]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/5.5/1, the various reagents are added in the amounts specified below and in the following order:

(284) TABLE-US-00029 Lyophilized compound A4 730 mg Lyophilized polyanionic compound 1 2000 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(285) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(286) The final pH is adjusted to 7.4±0.4.

(287) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(288) B39. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(289) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[lispro insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(290) TABLE-US-00030 Lyophilized substituted anionic compound A5 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL Sodium citrate solution at 1.188M 783 μL

(291) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(292) The final pH is adjusted to 7.4±0.4.

(293) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(294) B40. Preparation of a Solution of Human Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(295) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[human insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(296) TABLE-US-00031 Lyophilized substituted anionic compound A5 730 mg Commercial solution of Humulin ® R 100 mL Sodium citrate solution at 1.188M 783 μL

(297) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(298) The final pH is adjusted to 7.4±0.4.

(299) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(300) B41. Preparation of a Solution of Aspart Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(301) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[aspart insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(302) TABLE-US-00032 Lyophilized substituted anionic compound A5 730 mg Commercial solution of Novolog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(303) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(304) The final pH is adjusted to 7.4±0.4.

(305) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(306) B42. Preparation of a Solution of Glulisine Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(307) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[glulisine insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(308) TABLE-US-00033 Lyophilized substituted anionic compound A5 730 mg Commercial solution of Apidra ® 100 mL Sodium citrate solution at 1.188M 783 μL

(309) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(310) The final pH is adjusted to 7.4±0.4.

(311) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(312) B43. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A5 and of the Polyanionic Compound 1

(313) For a final volume of 100 mL of formulation, with a [compound A5]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/2.0/1, the various reagents are added in the amounts specified below and in the following order:

(314) TABLE-US-00034 Lyophilized compound A5 730 mg Lyophilized polyanionic compound 1 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(315) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(316) The final pH is adjusted to 7.4±0.4.

(317) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(318) B44. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A5 and of the Polyanionic Compound 1

(319) For a final volume of 100 mL of formulation, with a [compound A5]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are added in the amounts specified below:

(320) TABLE-US-00035 Compound A5 in lyophilized form 730 mg Polyanionic compound 1 in lyophilized form 1460 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(321) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(322) The final pH is adjusted to 7.4±0.4.

(323) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(324) B45. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A5 and of the Polyanionic Compound 1

(325) For a final volume of 100 mL of formulation, with a [compound A5]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/5.5/1, the various reagents are added in the amounts specified below and in the following order:

(326) TABLE-US-00036 Lyophilized compound A5 730 mg Lyophilized polyanionic compound 1 2000 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(327) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(328) The final pH is adjusted to 7.4±0.4.

(329) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(330) B47. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(331) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[lispro insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(332) TABLE-US-00037 Lyophilized substituted anionic compound A6 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL Sodium citrate solution at 1.188M 783 μL

(333) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(334) The final pH is adjusted to 7.4±0.4.

(335) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(336) B48. Preparation of a Solution of Human Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(337) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[human insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(338) TABLE-US-00038 Lyophilized substituted anionic compound A6 730 mg Commercial solution of Humulin ® R 100 mL Sodium citrate solution at 1.188M 783 μL

(339) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(340) The final pH is adjusted to 7.4±0.4.

(341) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(342) B49. Preparation of a Solution of Aspart Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(343) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[aspart insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(344) TABLE-US-00039 Lyophilized substituted anionic compound A6 730 mg Commercial solution of Novolog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(345) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(346) The final pH is adjusted to 7.4±0.4.

(347) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(348) B50. Preparation of a Solution of Glulisine Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(349) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[glulisine insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(350) TABLE-US-00040 Lyophilized substituted anionic compound A6 730 mg Commercial solution of Apidra ® 100 mL Sodium citrate solution at 1.188M 783 μL

(351) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(352) The final pH is adjusted to 7.4±0.4.

(353) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(354) B51. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A6 and of the Polyanionic Compound 1

(355) For a final volume of 100 mL of formulation, with a [compound A6]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/2.0/1, the various reagents are added in the amounts specified below and in the following order:

(356) TABLE-US-00041 Lyophilized compound A6 730 mg Lyophilized polyanionic compound 1 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(357) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(358) The final pH is adjusted to 7.4±0.4.

(359) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(360) B52. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A6 and of the Polyanionic Compound 1

(361) For a final volume of 100 mL of formulation, with a [compound A6]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are added in the amounts specified below:

(362) TABLE-US-00042 Compound A6 in lyophilized form 730 mg Polyanionic compound 1 in lyophilized form 1460 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(363) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(364) The final pH is adjusted to 7.4±0.4.

(365) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(366) B53. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A6 and of the Polyanionic Compound 1

(367) For a final volume of 100 mL of formulation, with a [compound A6]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/5.5/1, the various reagents are added in the amounts specified below and in the following order:

(368) TABLE-US-00043 Lyophilized compound A6 730 mg Lyophilized polyanionic compound 1 2000 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(369) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(370) The final pH is adjusted to 7.4±0.4.

(371) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(372) B55. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(373) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[lispro insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(374) TABLE-US-00044 Lyophilized substituted anionic compound A7 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL Sodium citrate solution at 1.188M 783 μL

(375) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(376) The final pH is adjusted to 7.4±0.4.

(377) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(378) B56. Preparation of a Solution of Human Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(379) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[human insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(380) TABLE-US-00045 Lyophilized substituted anionic compound A7 730 mg Commercial solution of Humulin ® R 100 mL Sodium citrate solution at 1.188M 783 μL

(381) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(382) The final pH is adjusted to 7.4±0.4.

(383) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(384) B57. Preparation of a Solution of Aspart Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(385) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[aspart insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(386) TABLE-US-00046 Lyophilized substituted anionic compound A7 730 mg Commercial solution of Novolog ® 100 mL Sodium citrate solution at 1.188M 783 μL

(387) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(388) The final pH is adjusted to 7.4±0.4.

(389) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(390) B58. Preparation of a Solution of Glulisine Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(391) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[glulisine insulin] mass ratio of 2.0 and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(392) TABLE-US-00047 Lyophilized substituted anionic compound A7 730 mg Commercial solution of Apidra ® 100 mL Sodium citrate solution at 1.188M 783 μL

(393) Citrate may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(394) The final pH is adjusted to 7.4±0.4.

(395) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(396) B59. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A7 and of the Polyanionic Compound 1

(397) For a final volume of 100 mL of formulation, with a [compound A7]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/2.0/1, the various reagents are added in the amounts specified below and in the following order:

(398) TABLE-US-00048 Lyophilized compound A7 730 mg Lyophilized polyanionic compound 1 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(399) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(400) The final pH is adjusted to 7.4±0.4.

(401) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(402) B60. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A7 and of the Polyanionic Compound 1

(403) For a final volume of 100 mL of formulation, with a [compound A7]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are added in the amounts specified below:

(404) TABLE-US-00049 Compound A7 in lyophilized form 730 mg Polyanionic compound 1 in lyophilized form 1460 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(405) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(406) The final pH is adjusted to 7.4±0.4.

(407) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(408) B61. Preparation of a Solution of Lispro Insulin at 100 IU/mL in the Presence of Compound A6 and of the Polyanionic Compound 1

(409) For a final volume of 100 mL of formulation, with a [compound A7]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2.0/5.5/1, the various reagents are added in the amounts specified below and in the following order:

(410) TABLE-US-00050 Lyophilized compound A7 730 mg Lyophilized polyanionic compound 1 2000 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL

(411) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(412) The final pH is adjusted to 7.4±0.4.

(413) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(414) B62. Preparation of a Solution of Insulin Analog (Lispro Insulin) at 200 IU/mL

(415) The commercial formulation of lispro insulin (Humalog®) was concentrated using Amicon Ultra-15 centrifugation tubes with a cut-off threshold of 3 kDa. The Amicon tubes were first rinsed with 12 mL of deionized water. 12 mL of the commercial formulation were centrifuged for 35 minutes at 4000 g at 20° C. The volume of the retentate was measured and the concentration thus determined. All the retentates were pooled and the global concentration was estimated (>200 IU/mL).

(416) The concentration of this concentrated lispro insulin solution was adjusted to 200 IU/mL by addition of the commercial formulation of lispro insulin (Humalog®). The concentrated lispro insulin formulation has the same concentrations of excipients (m-cresol, glycerol, phosphate) as the commercial formulation at 100 IU/mL.

(417) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(418) B63. Preparation of a Solution of Human Insulin at 200 IU/mL

(419) The commercial formulation of human insulin (Humulin® R) was concentrated using Amicon Ultra-15 centrifugation tubes with a cut-off threshold of 3 kDa. The Amicon tubes were first rinsed with 12 mL of deionized water. 12 mL of the commercial formulation were centrifuged for 35 minutes at 4000 g at 20° C. The volume of the retentate was measured and the concentration thus determined. All the retentates were pooled and the global concentration was estimated (>200 IU/mL).

(420) The concentration of this concentrated human insulin solution was adjusted to 200 IU/mL by addition of the commercial formulation of human insulin (Humulin® R). The concentrated human insulin formulation has the same concentrations of excipients (m-cresol, glycerol) as the commercial formulation at 100 IU/mL.

(421) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(422) B64. Preparation of a Solution of Aspart Insulin at 200 IU/mL

(423) The commercial formulation of aspart insulin (Novolog®) was concentrated using Amicon Ultra-15 centrifugation tubes with a cut-off threshold of 3 kDa. The Amicon tubes were first rinsed with 12 mL of deionized water. 12 mL of the commercial formulation were centrifuged for 35 minutes at 4000 g at 20° C. The volume of the retentate was measured and the concentration thus determined. All the retentates were pooled and the global concentration was estimated (>200 IU/mL).

(424) The concentration of this concentrated aspart insulin solution was adjusted to 200 IU/mL by addition of the commercial formulation of aspart insulin (Novolog®). The concentrated aspart insulin formulation has the same concentrations of excipients (m-cresol, glycerol) as the commercial formulation at 100 IU/mL.

(425) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(426) B65. Preparation of a Solution of Glulisine Insulin at 200 IU/mL

(427) The commercial formulation of glulisine insulin (Apidra®) was concentrated using Amicon Ultra-15 centrifugation tubes with a cut-off threshold of 3 kDa. The Amicon tubes were first rinsed with 12 mL of deionized water. 12 mL of the commercial formulation were centrifuged for 35 minutes at 4000 g at 20° C. The volume of the retentate was measured and the concentration thus determined. All the retentates were pooled and the global concentration was estimated (>200 IU/mL).

(428) The concentration of this concentrated glulisine insulin solution was adjusted to 200 IU/mL by addition of the commercial formulation of glulisine insulin (Apidra®). The concentrated glulisine insulin formulation has the same concentrations of excipients (m-cresol, NaCl, TRIS) as the commercial formulation at 100 IU/mL.

(429) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(430) B66. Preparation of a Solution of Human Insulin, of Lispro Insulin, of Aspart Insulin or of Glulisine Insulin at 300, 400 and 500 IU/mL

(431) Concentrated formulations of human insulin, of lispro insulin, of aspart insulin or of glulisine insulin at 300 IU/mL, 400 IU/mL or 500 IU/mL (and also at all intermediate concentrations) are prepared on the basis of the protocol of Example B65 relating to the preparation of a glulisine insulin solution at 200 IU/mL. The commercial insulin formulation is concentrated using Amicon Ultra-15 centrifugation tubes with a cut-off threshold of 3 kDa. The Amicon tubes are first rinsed with 12 mL of deionized water. 12 mL of the commercial formulation are centrifuged at 4000 g and at 20° C. By modifying the centrifugation time, it is possible to adjust the final concentration of insulin in the formulation. The volume of the retentate is measured and the concentration is thus estimated. All the retentates are pooled and the global concentration is estimated (>300, 400 or 500 IU/mL).

(432) The concentration of this concentrated insulin solution is adjusted to the desired concentration (e.g. 300 IU/mL, 400 IU/mL or 500 IU/mL) by addition of the insulin formulation (Humulin® R, Novolog®, Humalog® or Apidra®). The concentrated insulin formulation has the same concentrations of excipients as the commercial formulation at 100 IU/mL.

(433) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(434) B67. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(435) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[lispro insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(436) TABLE-US-00051 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 1460 mg Sodium citrate solution at 1.188M 1566 μL

(437) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(438) B68. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A1 and of the Polyanionic Compound 1

(439) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(440) TABLE-US-00052 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(441) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(442) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(443) B69. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A1 and of the Polyanionic Compound 1

(444) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(445) TABLE-US-00053 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(446) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(447) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(448) B70. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(449) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[human insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(450) TABLE-US-00054 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 1460 mg Sodium citrate solution at 1.188M 1566 μL

(451) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(452) B71. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A1 and of the Polyanionic Compound 1

(453) For a final volume of 100 mL of formulation with a [substituted anionic compound A1]/[polyanionic compound 1]/[human insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(454) TABLE-US-00055 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(455) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(456) B72. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A1 and of the Polyanionic Compound 1

(457) For a final volume of 100 mL of formulation with a [substituted anionic compound A1]/[polyanionic compound 1]/[human insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(458) TABLE-US-00056 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(459) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(460) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(461) B73. Preparation of a Solution of Aspart Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A1 at 14.6 mg/mL and 18.6 mM of Citrate

(462) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(463) TABLE-US-00057 Lyophilizate of substituted anionic compound A1 1460 mg Aspart insulin at 200 IU/mL 100 mL Sodium citrate solution at 1.188M 1566 μL

(464) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(465) B74. Preparation of a Solution of Glulisine Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(466) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[glulisine insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below and in the following order:

(467) TABLE-US-00058 Lyophilizate of substituted anionic compound A1 1460 mg Glulisine insulin at 200 IU/mL 100 mL Sodium citrate solution at 1.188M 1566 μL

(468) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(469) B75. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(470) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[lispro insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(471) TABLE-US-00059 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A2 1460 mg Sodium citrate solution at 1.188M 1566 μL

(472) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(473) B76. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A2 and of the Polyanionic Compound 1

(474) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(475) TABLE-US-00060 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A2 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(476) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(477) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(478) B77. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A2 and of the Polyanionic Compound 1

(479) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(480) TABLE-US-00061 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A2 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(481) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(482) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(483) B78. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(484) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[human insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(485) TABLE-US-00062 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A2 1460 mg Sodium citrate solution at 1.188M 1566 μL

(486) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(487) B79. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A2 and of the Polyanionic Compound 1

(488) For a final volume of 100 mL of formulation with a [substituted anionic compound A2]/[polyanionic compound 1]/[human insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(489) TABLE-US-00063 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A2 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(490) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(491) B80. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A2 and of the Polyanionic Compound 1

(492) For a final volume of 100 mL of formulation with a [substituted anionic compound A2]/[polyanionic compound 1]/[human insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(493) TABLE-US-00064 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A2 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(494) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(495) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(496) B81. Preparation of a Solution of Aspart Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(497) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below and in the following order:

(498) TABLE-US-00065 Lyophilizate of substituted anionic compound A2 1460 mg Aspart insulin at 200 IU/mL 100 mL Sodium citrate solution at 1.188M 1566 μL

(499) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(500) B82. Preparation of a Solution of Glulisine Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(501) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[glulisine insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below and in the following order:

(502) TABLE-US-00066 Lyophilizate of substituted anionic compound A2 1460 mg Glulisine insulin at 200 IU/mL 100 mL Sodium citrate solution at 1.188M 1566 μL

(503) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(504) B83. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(505) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[lispro insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(506) TABLE-US-00067 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 1460 mg Sodium citrate solution at 1.188M 1566 μL

(507) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(508) B84. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A3 and of the Polyanionic Compound 1

(509) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(510) TABLE-US-00068 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(511) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(512) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(513) B85. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A3 and of the Polyanionic Compound 1

(514) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(515) TABLE-US-00069 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(516) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(517) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(518) B86. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(519) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[human insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(520) TABLE-US-00070 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 1460 mg Sodium citrate solution at 1.188M 1566 μL

(521) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(522) B87. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A3 and of the Polyanionic Compound 1

(523) For a final volume of 100 mL of formulation with a [substituted anionic compound A3]/[polyanionic compound 1]/[human insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(524) TABLE-US-00071 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(525) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(526) B88. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A3 and of the Polyanionic Compound 1

(527) For a final volume of 100 mL of formulation with a [substituted anionic compound A3]/[polyanionic compound 1]/[human insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(528) TABLE-US-00072 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(529) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(530) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(531) B89. Preparation of a Solution of Aspart Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(532) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below and in the following order:

(533) TABLE-US-00073 Lyophilizate of substituted anionic compound A3 1460 mg Aspart insulin at 200 IU/mL 100 mL Sodium citrate solution at 1.188M 1566 μL

(534) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(535) B90. Preparation of a Solution of Glulisine Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(536) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[glulisine insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below and in the following order:

(537) TABLE-US-00074 Lyophilizate of substituted anionic compound A3 1460 mg Glulisine insulin at 200 IU/mL 100 mL Sodium citrate solution at 1.188M 1566 μL

(538) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(539) B91. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(540) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[lispro insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(541) TABLE-US-00075 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A4 1460 mg Sodium citrate solution at 1.188M 1566 μL

(542) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(543) B92. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A4 and of the Polyanionic Compound 1

(544) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(545) TABLE-US-00076 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A4 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(546) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(547) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(548) B93. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A4 and of the Polyanionic Compound 1

(549) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(550) TABLE-US-00077 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A4 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(551) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(552) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(553) B94. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(554) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[human insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(555) TABLE-US-00078 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A4 1460 mg Sodium citrate solution at 1.188M 1566 μL

(556) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(557) B95. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A4 and of the Polyanionic Compound 1

(558) For a final volume of 100 mL of formulation with a [substituted anionic compound A4]/[polyanionic compound 1]/[human insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(559) TABLE-US-00079 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A4 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(560) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(561) B96. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A4 and of the Polyanionic Compound 1

(562) For a final volume of 100 mL of formulation with a [substituted anionic compound A4]/[polyanionic compound 1]/[human insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(563) TABLE-US-00080 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A4 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(564) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(565) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(566) B97. Preparation of a Solution of Aspart Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(567) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below and in the following order:

(568) TABLE-US-00081 Lyophilizate of substituted anionic compound A4 1460 mg Aspart insulin at 200 IU/mL 100 mL Sodium citrate solution at 1.188M 1566 μL

(569) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(570) B98. Preparation of a Solution of Glulisine Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(571) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[glulisine insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below and in the following order:

(572) TABLE-US-00082 Lyophilizate of substituted anionic compound A4 1460 mg Glulisine insulin at 200 IU/mL 100 mL Sodium citrate solution at 1.188M 1566 μL

(573) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(574) B99. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(575) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[lispro insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(576) TABLE-US-00083 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A5 1460 mg Sodium citrate solution at 1.188M 1566 μL

(577) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(578) B100. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A5 and of the Polyanionic Compound 1

(579) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(580) TABLE-US-00084 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A5 1460 mg Lyophilizate of polyanionic compound 1 460 mg

(581) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(582) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(583) B101. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A5 and of the Polyanionic Compound 1

(584) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(585) TABLE-US-00085 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A5 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(586) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(587) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(588) B102. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(589) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[human insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(590) TABLE-US-00086 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A5 1460 mg Sodium citrate solution at 1.188M 1566 μL

(591) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(592) B103. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A5 and of the Polyanionic compound 1

(593) For a final volume of 100 mL of formulation with a [substituted anionic compound A5]/[polyanionic compound 1]/[human insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(594) TABLE-US-00087 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A5 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(595) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(596) B104. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A5 and of the Polyanionic Compound 1

(597) For a final volume of 100 mL of formulation with a [substituted anionic compound A5]/[polyanionic compound 1]/[human insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(598) TABLE-US-00088 Human insulin at 200 IU/mL  100 mL Lyophilizate of substituted anionic compound A5 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(599) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(600) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(601) B105. Preparation of a Solution of Aspart Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(602) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below and in the following order:

(603) TABLE-US-00089 Lyophilizate of substituted anionic compound A5 1460 mg Aspart insulin at 200 IU/mL  100 mL Sodium citrate solution at 1.188M 1566 μL

(604) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(605) B106. Preparation of a Solution of Glulisine Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(606) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[glulisine insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below and in the following order:

(607) TABLE-US-00090 Lyophilizate of substituted anionic compound A5 1460 mg Glulisine insulin at 200 IU/mL  100 mL Sodium citrate solution at 1.188M 1566 μL

(608) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(609) B107. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(610) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[lispro insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(611) TABLE-US-00091 Lispro insulin at 200 IU/mL  100 mL Lyophilizate of substituted anionic compound A6 1460 mg Sodium citrate solution at 1.188M 1566 μL

(612) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(613) B108. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A6 and of the Polyanionic Compound 1

(614) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(615) TABLE-US-00092 Lispro insulin at 200 IU/mL  100 mL Lyophilizate of substituted anionic compound A6 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(616) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(617) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(618) B109. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A6 and of the Polyanionic Compound 1

(619) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(620) TABLE-US-00093 Lispro insulin at 200 IU/mL  100 mL Lyophilizate of substituted anionic compound A6 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(621) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(622) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(623) B110. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(624) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[human insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(625) TABLE-US-00094 Human insulin at 200 IU/mL  100 mL Lyophilizate of substituted anionic compound A6 1460 mg Sodium citrate solution at 1.188M 1566 μL

(626) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(627) B111. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A6 and of the Polyanionic compound 1

(628) For a final volume of 100 mL of formulation with a [substituted anionic compound A6]/[polyanionic compound 1]/[human insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(629) TABLE-US-00095 Human insulin at 200 IU/mL  100 mL Lyophilizate of substituted anionic compound A6 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(630) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(631) B112. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A6 and of the Polyanionic compound 1

(632) For a final volume of 100 mL of formulation with a [substituted anionic compound A6]/[polyanionic compound 1]/[human insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(633) TABLE-US-00096 Human insulin at 200 IU/mL  100 mL Lyophilizate of substituted anionic compound A6 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(634) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(635) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(636) B113. Preparation of a Solution of Aspart Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(637) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below and in the following order:

(638) TABLE-US-00097 Lyophilizate of substituted anionic compound A6 1460 mg Aspart insulin at 200 IU/mL  100 mL Sodium citrate solution at 1.188M 1566 μL

(639) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(640) B114. Preparation of a Solution of Glulisine Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(641) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[glulisine insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below and in the following order:

(642) TABLE-US-00098 Lyophilizate of substituted anionic compound A6 1460 mg Glulisine insulin at 200 IU/mL  100 mL Sodium citrate solution at 1.188M 1566 μL

(643) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(644) B115. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(645) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[lispro insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(646) TABLE-US-00099 Lispro insulin at 200 IU/mL  100 mL Lyophilizate of substituted anionic compound A7 1460 mg Sodium citrate solution at 1.188M 1566 μL

(647) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(648) B116. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A7 and of the Polyanionic Compound 1

(649) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(650) TABLE-US-00100 Lispro insulin at 200 IU/mL  100 mL Lyophilizate of substituted anionic compound A7 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(651) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(652) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(653) B117. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A7 and of the Polyanionic Compound 1

(654) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[polyanionic compound 1]/[lispro insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(655) TABLE-US-00101 Lispro insulin at 200 IU/mL  100 mL Lyophilizate of substituted anionic compound A7 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(656) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(657) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(658) B118. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(659) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[human insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(660) TABLE-US-00102 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 1460 mg Sodium citrate solution at 1.188M 1566 μL

(661) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(662) B119. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A7 and of the Polyanionic Compound 1

(663) For a final volume of 100 mL of formulation with a [substituted anionic compound A7]/[polyanionic compound 1]/[human insulin] mass ratio of 2/2/1, the various reagents are mixed in the amounts specified below:

(664) TABLE-US-00103 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 1460 mg Lyophilizate of polyanionic compound 1 1460 mg

(665) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(666) B120. Preparation of a Solution of Human Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A7 and of the Polyanionic Compound 1

(667) For a final volume of 100 mL of formulation with a [substituted anionic compound A7]/[polyanionic compound 1]/[human insulin] mass ratio of 2/4/1, the various reagents are mixed in the amounts specified below:

(668) TABLE-US-00104 Human insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 1460 mg Lyophilizate of polyanionic compound 1 2920 mg

(669) The polyanionic compound 1 may be used in the acid form or in the basic form in the form of the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(670) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(671) B121. Preparation of a Solution of Aspart Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(672) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below and in the following order:

(673) TABLE-US-00105 Lyophilizate of substituted anionic compound A7 1460 mg Aspart insulin at 200 IU/mL 100 mL Sodium citrate solution at 1.188M 1566 μL

(674) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(675) B122. Preparation of a Solution of Glulisine Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(676) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[glulisine insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below and in the following order:

(677) TABLE-US-00106 Lyophilizate of substituted anionic compound A7 1460 mg Glulisine insulin at 200 IU/mL 100 mL Sodium citrate solution at 1.188M 1566 μL

(678) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(679) B123. Preparation of a Solution of Lispro Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(680) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(681) TABLE-US-00107 Lispro insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 2190 mg Sodium citrate 720 mg

(682) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(683) B124. Preparation of a Solution of Human Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(684) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(685) TABLE-US-00108 Human insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 2190 mg Sodium citrate 720 mg

(686) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(687) B125. Preparation of a Solution of Glulisine Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(688) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(689) TABLE-US-00109 Glulisine insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 2190 mg Sodium citrate 720 mg

(690) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(691) B126. Preparation of a Solution of Aspart Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(692) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(693) TABLE-US-00110 Aspart insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 2190 mg Sodium citrate 720 mg

(694) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(695) B127. Preparation of a Solution of Lispro Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(696) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(697) TABLE-US-00111 Lispro insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 2920 mg Sodium citrate 960 mg

(698) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(699) B128. Preparation of a Solution of Human Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(700) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(701) TABLE-US-00112 Human insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 2920 mg Sodium citrate 960 mg

(702) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(703) B129. Preparation of a Solution of Glulisine Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(704) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(705) TABLE-US-00113 Glulisine insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 2920 mg Sodium citrate 960 mg

(706) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(707) B130. Preparation of a Solution of Aspart Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(708) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[aspart insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(709) TABLE-US-00114 Aspart insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 2920 mg Sodium citrate 960 mg

(710) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(711) B131. Preparation of a Solution of Lispro Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(712) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(713) TABLE-US-00115 Lispro insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 3650 mg Sodium citrate 1200 mg

(714) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(715) B132. Preparation of a Solution of Human Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(716) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(717) TABLE-US-00116 Human insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 3650 mg Sodium citrate 1200 mg

(718) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(719) B133. Preparation of a Solution of Glulisine Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(720) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(721) TABLE-US-00117 Glulisine insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A1 3650 mg Sodium citrate 1200 mg

(722) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(723) B134. Preparation of a Solution of Aspart Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(724) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(725) TABLE-US-00118 Aspart insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A1 3650 mg Sodium citrate 1200 mg

(726) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(727) B135. Preparation of a Solution of Lispro Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(728) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(729) TABLE-US-00119 Lispro insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 2190 mg Sodium citrate  720 mg

(730) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(731) B136. Preparation of a Solution of Human Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(732) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(733) TABLE-US-00120 Human insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 2190 mg Sodium citrate  720 mg

(734) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(735) B137. Preparation of a Solution of Glulisine Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(736) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(737) TABLE-US-00121 Glulisine insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 2190 mg Sodium citrate  720 mg

(738) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(739) B138. Preparation of a Solution of Aspart Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(740) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(741) TABLE-US-00122 Aspart insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 2190 mg Sodium citrate  720 mg

(742) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(743) B139. Preparation of a Solution of Lispro Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(744) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(745) TABLE-US-00123 Lispro insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 2920 mg Sodium citrate  960 mg

(746) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(747) B140. Preparation of a Solution of Human Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(748) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(749) TABLE-US-00124 Human insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 2920 mg Sodium citrate  960 mg

(750) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(751) B141. Preparation of a Solution of Glulisine Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(752) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(753) TABLE-US-00125 Glulisine insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 2920 mg Sodium citrate  960 mg

(754) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(755) B142. Preparation of a Solution of Aspart Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(756) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[aspart insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(757) TABLE-US-00126 Aspart insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 2920 mg Sodium citrate  960 mg

(758) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(759) B143. Preparation of a Solution of Lispro Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(760) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(761) TABLE-US-00127 Lispro insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 3650 mg Sodium citrate 1200 mg

(762) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(763) B144. Preparation of a Solution of Human Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(764) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(765) TABLE-US-00128 Human insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 3650 mg Sodium citrate 1200 mg

(766) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(767) B145. Preparation of a Solution of Glulisine Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(768) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(769) TABLE-US-00129 Glulisine insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 3650 mg Sodium citrate 1200 mg

(770) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(771) B146. Preparation of a Solution of Aspart Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(772) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(773) TABLE-US-00130 Aspart insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A2 3650 mg Sodium citrate 1200 mg

(774) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(775) B147. Preparation of a Solution of Lispro Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(776) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(777) TABLE-US-00131 Lispro insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 2190 mg Sodium citrate  720 mg

(778) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(779) B148. Preparation of a Solution of Human Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(780) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(781) TABLE-US-00132 Human insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 2190 mg Sodium citrate  720 mg

(782) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(783) B149. Preparation of a Solution of Glulisine Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(784) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(785) TABLE-US-00133 Glulisine insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 2190 mg Sodium citrate  720 mg

(786) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(787) B150. Preparation of a Solution of Aspart Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(788) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(789) TABLE-US-00134 Aspart insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 2190 mg Sodium citrate  720 mg

(790) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(791) B151. Preparation of a Solution of Lispro Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(792) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(793) TABLE-US-00135 Lispro insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 2920 mg Sodium citrate  960 mg

(794) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(795) B152. Preparation of a Solution of Human Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(796) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(797) TABLE-US-00136 Human insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 2920 mg Sodium citrate  960 mg

(798) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(799) B153. Preparation of a Solution of Glulisine Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(800) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(801) TABLE-US-00137 Glulisine insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 2920 mg Sodium citrate  960 mg

(802) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(803) B154. Preparation of a Solution of Aspart Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(804) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[aspart insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(805) TABLE-US-00138 Aspart insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 2920 mg Sodium citrate  960 mg

(806) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(807) B155. Preparation of a Solution of Lispro Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(808) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(809) TABLE-US-00139 Lispro insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 3650 mg Sodium citrate 1200 mg

(810) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(811) B156. Preparation of a Solution of Human Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(812) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(813) TABLE-US-00140 Human insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 3650 mg Sodium citrate 1200 mg

(814) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(815) B157. Preparation of a Solution of Glulisine Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(816) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(817) TABLE-US-00141 Glulisine insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 3650 mg Sodium citrate 1200 mg

(818) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(819) B158. Preparation of a Solution of Aspart Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(820) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(821) TABLE-US-00142 Aspart insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 3650 mg Sodium citrate 1200 mg

(822) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(823) B159. Preparation of a Solution of Lispro Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(824) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(825) TABLE-US-00143 Lispro insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 2190 mg Sodium citrate  720 mg

(826) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(827) B160. Preparation of a Solution of Human Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(828) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(829) TABLE-US-00144 Human insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 2190 mg Sodium citrate  720 mg

(830) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(831) B161. Preparation of a Solution of Glulisine Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(832) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(833) TABLE-US-00145 Glulisine insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 2190 mg Sodium citrate  720 mg

(834) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(835) B162. Preparation of a Solution of Aspart Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(836) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(837) TABLE-US-00146 Aspart insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 2190 mg Sodium citrate  720 mg

(838) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(839) B163. Preparation of a Solution of Lispro Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(840) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(841) TABLE-US-00147 Lispro insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 2920 mg Sodium citrate  960 mg

(842) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(843) B164. Preparation of a Solution of Human Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(844) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(845) TABLE-US-00148 Human insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 2920 mg Sodium citrate  960 mg

(846) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(847) B165. Preparation of a Solution of Glulisine Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(848) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(849) TABLE-US-00149 Glulisine insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 2920 mg Sodium citrate  960 mg

(850) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(851) B166. Preparation of a Solution of Aspart Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(852) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[aspart insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(853) TABLE-US-00150 Aspart insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 2920 mg Sodium citrate  960 mg

(854) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(855) B167. Preparation of a Solution of Lispro Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(856) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(857) TABLE-US-00151 Lispro insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 3650 mg Sodium citrate 1200 mg

(858) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(859) B168. Preparation of a Solution of Human Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(860) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(861) TABLE-US-00152 Human insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 3650 mg Sodium citrate 1200 mg

(862) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(863) B169. Preparation of a Solution of Glulisine Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(864) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(865) TABLE-US-00153 Glulisine insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A4 3650 mg Sodium citrate 1200 mg

(866) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(867) B170. Preparation of a solution of aspart insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(868) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(869) TABLE-US-00154 Aspart insulin at 500 IU/mL  100 mL Lyophilizate of substituted anionic compound A3 3650 mg Sodium citrate 1200 mg

(870) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(871) B171. Preparation of a Solution of Lispro Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(872) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(873) TABLE-US-00155 Lispro insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A5 2190 mg Sodium citrate  720 mg

(874) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(875) B172. Preparation of a Solution of Human Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(876) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(877) TABLE-US-00156 Human insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A5 2190 mg Sodium citrate  720 mg

(878) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(879) B173. Preparation of a Solution of Glulisine Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(880) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(881) TABLE-US-00157 Glulisine insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A5 2190 mg Sodium citrate  720 mg

(882) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(883) B174. Preparation of a Solution of Aspart Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(884) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(885) TABLE-US-00158 Aspart insulin at 300 IU/mL  100 mL Lyophilizate of substituted anionic compound A5 2190 mg Sodium citrate  720 mg

(886) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(887) B175. Preparation of a Solution of Lispro Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(888) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(889) TABLE-US-00159 Lispro insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A5 2920 mg Sodium citrate  960 mg

(890) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(891) B176. Preparation of a Solution of Human Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(892) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(893) TABLE-US-00160 Human insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A5 2920 mg Sodium citrate  960 mg

(894) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(895) B177. Preparation of a Solution of Glulisine Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(896) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(897) TABLE-US-00161 Glulisine insulin at 400 IU/mL  100 mL Lyophilizate of substituted anionic compound A5 2920 mg Sodium citrate  960 mg

(898) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(899) B178. Preparation of a Solution of Aspart Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(900) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[aspart insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(901) TABLE-US-00162 Aspart insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A5 2920 mg Sodium citrate 960 mg

(902) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(903) B179. Preparation of a Solution of Lispro Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(904) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(905) TABLE-US-00163 Lispro insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A5 3650 mg Sodium citrate 1200 mg

(906) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(907) B180. Preparation of a Solution of Human Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(908) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(909) TABLE-US-00164 Human insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A5 3650 mg Sodium citrate 1200 mg

(910) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(911) B181. Preparation of a Solution of Glulisine Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(912) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(913) TABLE-US-00165 Glulisine insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A5 3650 mg Sodium citrate 1200 mg

(914) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(915) B182. Preparation of a Solution of Aspart Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(916) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(917) TABLE-US-00166 Aspart insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 3650 mg Sodium citrate 1200 mg

(918) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(919) B183. Preparation of a Solution of Lispro Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(920) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(921) TABLE-US-00167 Lispro insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 2190 mg Sodium citrate 720 mg

(922) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(923) B184. Preparation of a Solution of Human Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(924) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(925) TABLE-US-00168 Human insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 2190 mg Sodium citrate 720 mg

(926) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(927) B185. Preparation of a Solution of Glulisine Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(928) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(929) TABLE-US-00169 Glulisine insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 2190 mg Sodium citrate 720 mg

(930) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(931) B186. Preparation of a Solution of Aspart Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(932) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(933) TABLE-US-00170 Aspart insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 2190 mg Sodium citrate 720 mg

(934) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(935) B187. Preparation of a Solution of Lispro Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(936) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(937) TABLE-US-00171 Lispro insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 2920 mg Sodium citrate 960 mg

(938) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(939) B188. Preparation of a Solution of Human Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(940) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(941) TABLE-US-00172 Human insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 2920 mg Sodium citrate 960 mg

(942) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(943) B189. Preparation of a Solution of Glulisine Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(944) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(945) TABLE-US-00173 Glulisine insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 2920 mg Sodium citrate 960 mg

(946) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(947) B190. Preparation of a Solution of Aspart Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(948) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[aspart insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(949) TABLE-US-00174 Aspart insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 2920 mg Sodium citrate 960 mg

(950) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(951) B191. Preparation of a Solution of Lispro Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(952) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(953) TABLE-US-00175 Lispro insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 3650 mg Sodium citrate 1200 mg

(954) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(955) B192. Preparation of a Solution of Human Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(956) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(957) TABLE-US-00176 Human insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 3650 mg Sodium citrate 1200 mg

(958) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(959) B193. Preparation of a Solution of Glulisine Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(960) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(961) TABLE-US-00177 Glulisine insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 3650 mg Sodium citrate 1200 mg

(962) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(963) B194. Preparation of a Solution of Aspart Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(964) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(965) TABLE-US-00178 Aspart insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 3650 mg Sodium citrate 1200 mg

(966) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(967) B195. Preparation of a Solution of Lispro Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(968) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(969) TABLE-US-00179 Lispro insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 2190 mg Sodium citrate 720 mg

(970) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(971) B196. Preparation of a Solution of Human Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(972) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(973) TABLE-US-00180 Human insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 2190 mg Sodium citrate 720 mg

(974) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(975) B197. Preparation of a Solution of Glulisine Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(976) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(977) TABLE-US-00181 Glulisine insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 2190 mg Sodium citrate 720 mg

(978) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(979) B198. Preparation of a Solution of Aspart Insulin at 300 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(980) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(981) TABLE-US-00182 Aspart insulin at 300 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 2190 mg Sodium citrate 720 mg

(982) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(983) B199. Preparation of a Solution of Lispro Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(984) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(985) TABLE-US-00183 Lispro insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 2920 mg Sodium citrate 960 mg

(986) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(987) B200. Preparation of a Solution of Human Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(988) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(989) TABLE-US-00184 Human insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 2920 mg Sodium citrate 960 mg

(990) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(991) B201. Preparation of a Solution of Glulisine Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(992) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(993) TABLE-US-00185 Glulisine insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 2920 mg Sodium citrate 960 mg

(994) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(995) B202. Preparation of a Solution of Aspart Insulin at 400 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(996) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[aspart insulin] mass ratio of 2, the various reagents are mixed in the amounts specified below:

(997) TABLE-US-00186 Aspart insulin at 400 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 2920 mg Sodium citrate 960 mg

(998) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(999) B203. Preparation of a Solution of Lispro Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(1000) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[lispro insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(1001) TABLE-US-00187 Lispro insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 3650 mg Sodium citrate 1200 mg

(1002) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1003) B204. Preparation of a Solution of Human Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(1004) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[human insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(1005) TABLE-US-00188 Human insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 3650 mg Sodium citrate 1200 mg

(1006) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1007) B205. Preparation of a Solution of Glulisine Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(1008) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[glulisine insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(1009) TABLE-US-00189 Glulisine insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 3650 mg Sodium citrate 1200 mg

(1010) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1011) B206. Preparation of a Solution of Aspart Insulin at 500 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(1012) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[aspart insulin] mass ratio of 2.0, the various reagents are mixed in the amounts specified below:

(1013) TABLE-US-00190 Aspart insulin at 500 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 3650 mg Sodium citrate 1200 mg

(1014) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1015) B207. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A1 and Citrate

(1016) For a final volume of 100 mL of formulation, with a [substituted anionic compound A1]/[lispro insulin] mass ratio of 1, the various reagents are mixed in the amounts specified below:

(1017) TABLE-US-00191 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A1 730 mg Sodium citrate solution at 1.188M 783 μL

(1018) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1019) B208. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A2 and Citrate

(1020) For a final volume of 100 mL of formulation, with a [substituted anionic compound A2]/[lispro insulin] mass ratio of 1, the various reagents are mixed in the amounts specified below:

(1021) TABLE-US-00192 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A2 730 mg Sodium citrate solution at 1.188M 783 μL

(1022) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1023) B209. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A3 and Citrate

(1024) For a final volume of 100 mL of formulation, with a [substituted anionic compound A3]/[lispro insulin] mass ratio of 1, the various reagents are mixed in the amounts specified below:

(1025) TABLE-US-00193 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A3 730 mg Sodium citrate solution at 1.188M 783 μL

(1026) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1027) B210. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A4 and Citrate

(1028) For a final volume of 100 mL of formulation, with a [substituted anionic compound A4]/[lispro insulin] mass ratio of 1, the various reagents are mixed in the amounts specified below:

(1029) TABLE-US-00194 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A4 730 mg Sodium citrate solution at 1.188M 783 μL

(1030) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1031) B211. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A5 and Citrate

(1032) For a final volume of 100 mL of formulation, with a [substituted anionic compound A5]/[lispro insulin] mass ratio of 1, the various reagents are mixed in the amounts specified below:

(1033) TABLE-US-00195 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A5 730 mg Sodium citrate solution at 1.188M 783 μL

(1034) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1035) B212. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A6 and Citrate

(1036) For a final volume of 100 mL of formulation, with a [substituted anionic compound A6]/[lispro insulin] mass ratio of 1, the various reagents are mixed in the amounts specified below:

(1037) TABLE-US-00196 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A6 730 mg Sodium citrate solution at 1.188M 783 μL

(1038) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1039) B213. Preparation of a Solution of Lispro Insulin at 200 IU/mL in the Presence of the Substituted Anionic Compound A7 and Citrate

(1040) For a final volume of 100 mL of formulation, with a [substituted anionic compound A7]/[lispro insulin] mass ratio of 1, the various reagents are mixed in the amounts specified below:

(1041) TABLE-US-00197 Lispro insulin at 200 IU/mL 100 mL Lyophilizate of substituted anionic compound A7 730 mg Sodium citrate solution at 1.188M 783 μL

(1042) The final pH is adjusted to 7.4±0.4. The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1043) C Pharmacodynamics and Pharmacokinetics

(1044) Protocol for Measuring the Pharmacodynamics of the Insulin Solutions

(1045) Domestic pigs weighing about 50 kg, precatheterized in the jugular vein, are fasted for 2.5 hours before the start of the experiment. Within the hour prior to the injection of insulin, three blood samples are collected so as to determine the basal level of glucose and insulin.

(1046) The injection of insulin at a dose of 0.09 IU/kg for lispro insulin is performed subcutaneously in the neck, under the animal's ear, using an insulin pen (Novo, Sanofi or Lilly) equipped with a 31 G needle.

(1047) Blood samples are then collected every 4 minutes for 20 minutes and then every 10 minutes up to 3 hours. After each sample collection, the catheter is rinsed with a dilute heparin solution.

(1048) A drop of blood is collected to determine the glycemia using a glucometer.

(1049) The pharmacodynamic curves for glucose expressed as a percentage of the basal level are then plotted. The time required to reach the minimum glucose level in the blood and 50% of the minimum glucose level in the blood for each pig are determined and reported as Tmin glucose and T50% Rmin glucose, respectively. The mean values of the Tmin glucose and of the T50% Rmin glucose are then calculated.

(1050) The remaining blood is collected in a dry tube and is centrifuged to isolate the serum. The insulin levels in the serum samples are measured via the sandwich ELISA immunoenzymatic method for each pig.

(1051) The pharmacokinetic curves expressed as the delta of the basal level are then plotted. The time required to reach the maximum concentration and the time required to reach 50% of the maximum insulin concentration in the serum for each pig are determined and reported as Tmax insulin and T50% Cmax insulin, respectively. The mean values of the Tmax insulin and of the T50% Cmax insulin are then calculated.

(1052) C2: Pharmacodynamic and Pharmacokinetic Results for the Insulin Solutions of Examples B2 and B12

(1053) TABLE-US-00198 Substituted anionic Number of Example Insulin compound Excipient pigs B2 lispro — — 10 B12 lispro A2 Citrate 9.3 mM 11

(1054) The pharmacodynamic results obtained with the compositions described in examples B2 and B12 are presented in FIG. 1. Analysis of these curves shows that the composition of example B12 comprising the substituted anionic compound A2 and citrate as excipient (curve plotted with the squares corresponding to Example B12, Tmin glucose=42±12 min and T50% Rmin glucose=18±4 min) makes it possible to obtain faster action than that of the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmin glucose=47±11 min and T50% Rmin glucose=26±8 min).

(1055) The pharmacokinetic results obtained with the compositions described in examples B2 and B12 are presented in FIG. 2. Analysis of these curves shows that the composition of example B12 comprising the substituted anionic compound A2 and citrate as excipient (curve plotted with the squares corresponding to Example B12, Tmax insulin=24±18 min and T50% Cmax insulin=6±3 min) induces more rapid absorption of the lispro insulin than the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmax insulin=23±23 min and T50% Cmax insulin=17±22 min).

(1056) C3: Pharmacodynamic and Pharmacokinetic Results for the Insulin Solutions of Examples B2 and B7

(1057) TABLE-US-00199 Substituted anionic Number of Example Insulin compound Excipient pigs B2 lispro — — 10 B7 lispro A1 Citrate 9.3 mM 12

(1058) The pharmacodynamic results obtained with the compositions described in examples B2 and B7 are presented in FIG. 3. Analysis of these curves shows that the composition of example B7 comprising the substituted anionic compound A1 and citrate as excipient (curve plotted with the squares corresponding to Example B7, Tmin glucose=48±22 min and T50% Rmin glucose=17±7 min) makes it possible to obtain faster action than that of the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmin glucose=47±11 min and T50% Rmin glucose=26±8 min).

(1059) The pharmacokinetic results obtained with the compositions described in examples B2 and B7 are presented in FIG. 4. Analysis of these curves shows that the composition of example B7 comprising the substituted anionic compound A1 and citrate as excipient (curve plotted with the squares corresponding to Example B7, Tmax insulin=20±14 min and T50% Cmax insulin=6±3 min) induces more rapid absorption of the lispro insulin than the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmax insulin=23±23 min and T50% Cmax insulin=17±22 min).

(1060) C3′: Pharmacodynamic and Pharmacokinetic Results for the Insulin Solutions of Examples B2 and B7

(1061) TABLE-US-00200 Substituted anionic Number of Example Insulin compound Excipient pigs B2 lispro — —  9 B7 lispro A1 Citrate 9.3 mM 11

(1062) The pharmacodynamic results obtained with the compositions described in examples B2 and B7 are presented in FIG. 14. Analysis of these curves shows that the composition of example B7 comprising the substituted anionic compound A1 and citrate as excipient (curve plotted with the squares corresponding to Example B7, Tmin glucose=35±10 min and T50% Rmin glucose=14±4 min) makes it possible to obtain faster action than that of the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmin glucose=38±8 min and T50% Rmin glucose=18±10 min).

(1063) The pharmacokinetic results obtained with the compositions described in examples B2 and B7 are presented in FIG. 15. Analysis of these curves shows that the composition of example B7 comprising the substituted anionic compound A1 and citrate as excipient (curve plotted with the squares corresponding to Example B7, Tmax insulin=13±6 min and T50% Cmax insulin=5±4 min) induces more rapid absorption of the lispro insulin than the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmax insulin=19±8 min and T50% Cmax insulin=8±4 min).

(1064) C4: Pharmacodynamic and Pharmacokinetic Results for the Insulin Solutions of Examples B2 and B39

(1065) TABLE-US-00201 Substituted anionic Number of Example Insulin compound Excipient pigs B2 lispro — — 13 B39 lispro A5 Citrate 9.3 mM  8

(1066) The pharmacodynamic results obtained with the compositions described in examples B2 and B39 are presented in FIG. 7. Analysis of these curves shows that the composition of example B39 comprising the substituted anionic compound A5 and citrate as excipient (curve plotted with the squares corresponding to Example B39, Tmin glucose=51±25 min and T50% Rmin glucose=15±7 min) makes it possible to obtain faster action than that of the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmin glucose=46±18 min and T50% Rmin glucose=21±6 min).

(1067) The pharmacokinetic results obtained with the compositions described in examples B2 and B39 are presented in FIG. 8. Analysis of these curves shows that the composition of example B39 comprising the substituted anionic compound A5 and citrate as excipient (curve plotted with the squares corresponding to Example B39, Tmax insulin=13±12 min and T50% Cmax insulin=4±3 min) induces more rapid absorption of the lispro insulin than the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmax insulin=22±20 min and T50% Cmax insulin=7±4 min).

(1068) C5: Pharmacodynamic and Pharmacokinetic Results for the Insulin Solutions of Examples B2 and B47

(1069) TABLE-US-00202 Substituted anionic Number of Example Insulin compound Excipient pigs B2 lispro — — 13 B47 lispro A6 Citrate 9.3 mM 11

(1070) The pharmacodynamic results obtained with the compositions described in examples B2 and B47 are presented in FIG. 9. Analysis of these curves shows that the composition of example B47 comprising the substituted anionic compound A6 and citrate as excipient (curve plotted with the squares corresponding to Example B47, Tmin glucose=34±10 min and T50% Rmin glucose=13±3 min) makes it possible to obtain faster action than that of the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmin glucose=46±18 min and T50% Rmin glucose=21±6 min).

(1071) The pharmacokinetic results obtained with the compositions described in examples B2 and B47 are presented in FIG. 10. Analysis of these curves shows that the composition of example B47 comprising the substituted anionic compound A6 and citrate as excipient (curve plotted with the squares corresponding to Example B47, Tmax insulin=11±5 min and T50% Cmax insulin=5±3 min) induces more rapid absorption of the lispro insulin than the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmax insulin=22±20 min and T50% Cmax insulin=7±4 min).

(1072) C6: Pharmacodynamic and Pharmacokinetic Results for the Insulin Solutions of Examples B2 and B55

(1073) TABLE-US-00203 Substituted anionic Number of Example Insulin compound Excipient pigs B2 lispro — — 13 B55 lispro A7 Citrate 9.3 mM 12

(1074) The pharmacodynamic results obtained with the compositions described in examples B2 and B55 are presented in FIG. 11. Analysis of these curves shows that the composition of example B55 comprising the substituted anionic compound A7 and citrate as excipient (curve plotted with the squares corresponding to Example B55, Tmin glucose=34±15 min and T50% Rmin glucose=15±4 min) makes it possible to obtain faster action than that of the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmin glucose=46±18 min and T50% Rmin glucose=21±6 min).

(1075) The pharmacokinetic results obtained with the compositions described in examples B2 and B55 are presented in FIG. 12. Analysis of these curves shows that the composition of example B55 comprising the substituted anionic compound A7 and citrate as excipient (curve plotted with the squares corresponding to Example B55, Tmax insulin=13±9 min and T50% Cmax insulin=4±3 min) induces more rapid absorption of the lispro insulin than the Humalog® commercial composition of example B2 (curve plotted with the triangles corresponding to example B2, Tmax insulin=22±20 min and T50% Cmax insulin=7±4 min).

(1076) D Circular Dichroism (CD)

(1077) Circular dichroism makes it possible to study the secondary and quaternary structure of insulin. Insulin monomers become organized as dimers and as hexamers. The hexamer is the physically and chemically most stable form of insulin. The CD signal at 276 nm is characteristic of the hexameric form of insulin (hexamer signal at about −300°, dimer signal between −200° and −250° and monomer signal below −200°). Loss of the CD signal at 276 nm is thus characteristic of a destabilization of the hexamer into dimers or monomers.

(1078) D1. Impact of the Substituted Anionic Compound A1 on the CD Signal of Human Insulin at 276 nm

(1079) The results obtained are presented in FIG. 5. This figure describes on the y-axis the CD signal at 276 nm (deg.Math.cm.sup.2.Math.dmol.sup.−1) and, on the x-axis:

(1080) A: rhINS (human insulin) (100 IU/ml)

(1081) B: rhINS/EDTA

(1082) C: rhINS/Citrate

(1083) D: rhINS/EDTA/Citrate

(1084) E: rhINS/Compound A1 (100 IU/ml/7.3 mg/mL)

(1085) EDTA and the EDTA/citrate combination have a very pronounced impact on the hexameric structure of human insulin (total association of the hexamer into dimers). In contrast, citrate and the substituted anionic compound A1 have no impact on the hexameric structure of human insulin. In contrast with EDTA, the compositions based on the substituted anionic compound A1 do not dissociate the hexamer of human insulin.

(1086) D2. Impact of the Substituted Anionic Compounds A2 and A3 on the CD Signal of Human Insulin at 276 nm

(1087) The results obtained are presented in FIG. 6. This figure describes on the y-axis the CD signal at 276 nm (deg.Math.cm.sup.2.Math.dmol.sup.−1) and, on the x-axis:

(1088) F: rhINS (100 IU/ml)

(1089) G: rhINS/Compound A2 (100 IU/ml/7.3 mg/mL)

(1090) H: rhINS/Compound A3 (100 IU/ml/7.3 mg/mL)

(1091) The substituted anionic compounds A2 and A3 have no impact on the hexameric structure of human insulin. In contrast with EDTA, the compositions based on the substituted anionic compounds A2 and A3 do not dissociate the hexamer of human insulin.

(1092) D3 State of Association of Lispro Insulin Evaluated by Circular Dichroism in the Presence of Various Substituted Anionic Compounds and Citrate

(1093) Circular dichroism makes it possible to study the secondary and quaternary structure of insulin. Insulin monomers become organized as dimers and as hexamers. The hexamer is the physically and chemically most stable form of insulin. Two hexameric forms exist, the form R6 and the form T6. Lispro insulin has a strong signal at 240 nm characteristic of the hexameric form R6 (the most stable form). Loss of the signal at 240 nm is linked to a destabilization of the hexamer R6.

(1094) Preparation of a 1.010 M Sodium Citrate Solution

(1095) A sodium citrate solution is obtained by dissolving 14.9077 g of sodium citrate (50.69 mmol) in 50 mL of water in a graduated flask. The pH is adjusted to 7.4 by adding 0.21 mL of 1 M HCl.

(1096) Preparation of Solutions of Lispro Insulin at 100 IU/mL in the Presence of the Substituted Anionic Compound and Citrate

(1097) For a final volume of 100 mL of formulation, with a concentration of substituted anionic compound of 7.3 mg/mL and a concentration of 9.3 mM of citrate, the various reagents are added in the amounts specified below and in the following order:

(1098) TABLE-US-00204 Lyophilized substituted anionic compound 730 mg Commercial solution of Humalog ® at 100 IU/mL 100 mL Sodium citrate solution at 1.010M 921 μL

(1099) Citrate may be used in the acid form or in the basic form as the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(1100) The final pH is adjusted to 7.4±0.4.

(1101) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1102) Preparation of the Solution of Lispro Insulin at 100 IU/mL in the Presence of Citrate

(1103) For a final volume of 100 mL of formulation, with a concentration of 9.3 mM of citrate, the various compounds are added in the amounts specified below and in the following order:

(1104) TABLE-US-00205 Commercial solution of Humalog ® at 100 IU/mL 100 mL Sodium citrate solution at 1.010M 921 μL

(1105) Citrate may be used in the acid form or in the basic form as the sodium salt, the potassium salt or another salt that is compatible with an injectable formulation.

(1106) The final pH is adjusted to 7.4±0.4.

(1107) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1108) Preparation of the Solution of Lispro Insulin at 100 IU/mL in the Presence of EDTA

(1109) For a final volume of 100 mL of formulation, with a concentration of 0.3 mM of EDTA, the various compounds are added in the amounts specified below and in the following order:

(1110) TABLE-US-00206 Commercial solution of Humalog ® at 100 IU/mL 100 mL Commercial solution of EDTA at 0.5M  60 μL

(1111) The final pH is adjusted to 7.4±0.4.

(1112) The clear solution is filtered through a 0.22 μm membrane and stored at 4° C.

(1113) The results obtained are presented in FIG. 13. This figure describes on the y-axis the CD signal at 240 nm (deg.Math.cm.sup.2.Math.dmol.sup.−1) and, on the x-axis:

(1114) A: lispro insulin 100 IU/mL

(1115) B: lispro insulin+7.3 mg/mL of substituted anionic compound A1+citrate at 9.3 mM

(1116) C: lispro insulin+7.3 mg/mL of substituted anionic compound A2+citrate at 9.3 mM

(1117) D: lispro insulin+7.3 mg/mL of substituted anionic compound A5+citrate at 9.3 mM

(1118) E: lispro insulin+7.3 mg/mL of substituted anionic compound A6+citrate at 9.3 mM

(1119) F: lispro insulin+7.3 mg/mL of substituted anionic compound A7+citrate at 9.3 mM

(1120) G: lispro insulin+EDTA 300 μM

(1121) H: lispro insulin+citrate at 9.3 mM

(1122) EDTA completely destructures the R6 form of lispro insulin. EDTA thus has a pronounced effect on the hexamer. In contrast, citrate alone and the mixture of substituted anionic compound/citrate have no impact on the CD signal at 240 nm. These compounds thus have no impact on the R6 structure of the hexamer and, all the less so, on the hexameric structure.