Polyalkoxy fatty compound

Abstract

Provided is a composition comprising one or more protein and one or more polyalkoxy fatty compound and wherein said polyalkoxy fatty compound has structure (I) ##STR00001##
wherein R.sup.1 is a fatty group; R.sup.2 is H or a substituted or unsubstituted hydrocarbyl group; n is 0 to 5; X.sup.1 is O, S, or NH; X.sup.2 is O, S, or NH; and R.sup.3 is a polymeric group comprising polymerized units of (II) and (III) ##STR00002##

Claims

1. A composition comprising water, one or more protein, and one or more polyalkoxy fatty compound that has structure (I) ##STR00016## wherein R.sup.1 is a fatty group; R.sup.2 is H or a substituted or unsubstituted hydrocarbyl group; n is 0 to 5; X.sup.1 is O, S, or NH; X.sup.2 is O, S, or NH; R.sup.3 has the structure R.sup.4CH.sub.3, where R.sup.4 is a polymeric group consisting of polymerized units of (II) and (III); and R.sup.3 has number-average molecular weight of 600 or higher ##STR00017##

2. The composition of claim 1, wherein the polyalkoxy fatty compound is present in an amount of 0.01 mg/mL or more.

3. The composition of claim 1, wherein the protein is present in an amount of 400 mg/mL or less.

4. The composition of claim 1, wherein the water is present in an amount of 50% or more, by weight based on the weight of said composition, wherein said protein and said polyalkoxy fatty compound are dissolved or dispersed in said water.

5. The composition of claim 1, wherein the water is present in an amount of 50% or more, by weight based on the weight of said composition, wherein said protein and said polyalkoxy fatty compound are dissolved in said water.

6. The composition of claim 1, wherein said composition further comprises one or more additional ingredients, wherein the total amount of all additional ingredients is 300 mg/mL or less.

7. The composition of claim 1, wherein said protein is selected from monoclonal antibodies, growth factors, insulins, immunoglobulins, polyclonal antibodies, hormones, enzymes, polypeptides, fusions of peptides, glycosylated proteins, antigens, antigen subunits, and combinations thereof.

8. The composition of claim 1, wherein R.sup.1 is a linear unsubstituted alkyl group having 10 to 16 carbon atoms, wherein R.sup.2 is selected from the group consisting of hydrogen, methyl, and CH.sub.2(C.sub.6H.sub.5), wherein (C.sub.6H.sub.5) is a benzene ring, and wherein R.sup.3 has number-average molecular weight from 800 to 3000.

9. The composition of claim 1, wherein X.sup.2 is NH.

10. The composition of claim 1, wherein n is 0 or 1.

11. The composition of claim 1, wherein X.sup.1 is NH.

12. A composition comprising water, one or more protein, and one or more polyalkoxy fatty compound that has structure (I) ##STR00018## wherein R.sup.1 is a fatty group; R.sup.2 is H or a substituted or unsubstituted hydrocarbyl group; n is 1 to 5; X.sup.1 is O, S, or NH; X.sup.2 is O, S, or NH; and R.sup.3 is a polymeric group comprising polymerized units of (II) and (III) ##STR00019##

13. The composition of claim 12, wherein the polyalkoxy fatty compound is present in an amount of 0.01 mg/mL or more.

14. The composition of claim 12, wherein the protein is present in an amount of 400 mg/mL or less.

15. The composition of claim 12, wherein the water is present in an amount of 50% or more, by weight based on the weight of said composition, wherein said protein and said polyalkoxy fatty compound are dissolved or dispersed in said water.

16. The composition of claim 12, wherein the water is present in an amount of 50% or more, by weight based on the weight of said composition, wherein said protein and said polyalkoxy fatty compound are dissolved in said water.

17. The composition of claim 12, wherein said composition further comprises one or more additional ingredients, wherein the total amount of all additional ingredients is 300 mg/mL or less.

18. The composition of claim 12, wherein said protein is selected from monoclonal antibodies, growth factors, insulins, immunoglobulins, polyclonal antibodies, hormones, enzymes, polypeptides, fusions of peptides, glycosylated proteins, antigens, antigen subunits, and combinations thereof.

19. The composition of claim 12, wherein R.sup.1 is a linear unsubstituted alkyl group having 10 to 16 carbon atoms, wherein R.sup.2 is selected from the group consisting of hydrogen, methyl, and CH.sub.2(C.sub.6H.sub.5), wherein (C.sub.6H.sub.5) is a benzene ring, and wherein R.sup.3 has number-average molecular weight from 800 to 3000.

20. The composition of claim 12, wherein X.sup.2 is NH.

21. The composition of claim 12, wherein n is 1.

22. The composition of claim 12, wherein X.sup.1 is NH.

Description

EXAMPLE 1

Testing of Protein Solutions Containing BSA

(1) Scanning DLS of 5 mg/mL BSA with various surfactants as indicated. Scan speed was 0.05 C./min from 25 to 80 C. Surfactants were all 1 mg/mL. In scanning DLS, samples were tested for volume-average particle diameter as a function of temperature. Each sample eventually showed an increase in diameter. The temperature of the onset (T.sub.onset) was recorded. Higher T.sub.onset means a more stable protein solution.

(2) Nano-DSC of BSA (5 mg/mL) was performed with various surfactants (5 mg/mL). The Nano-DSC graph of heat flow vs. temperature shows a peak between 60 C. and 85 C. that is labeled the unfolding peak, and it indicates the temperature at which the protein denatures at maximal rate. The temperature of the unfolding peak (Tm) is reported. Higher Tm means a more stable the protein solution.

(3) Circular dichroism (CD) BSA samples (5 mg/mL) with different surfactants were measured following heat aging at 70 C. for 30 minutes. The graph of CD vs. wavelength shows a peak at 222 nm, indicating absorbance of the alpha-helix. Decrease in the alpha-helix peak indicates denaturation. For each sample, the CD at 222 nm was examined, and the difference between the value of the initial sample was compared to the value of the same sample after heat aging at 70 C. for 30 minutes; this difference is reported as CD. Higher absolute value of CD means a less stable protein solution.

(4) Also performed was isothermal DLS of BSA (5 mg/mL) with 1 mg/mL surfactants at 62 C. Volume-average particle diameter was measured as a function of time. The diameter at 2000 seconds (D2 k) was recorded. Lower value of D2 k means a more stable protein solution.

(5) Results were as follows. nt means not tested.

(6) TABLE-US-00002 T.sub.onset Tm CD D2k Type Surfactant ( C.) ( C.) (mdeg) (nm) comparative none 57 61 30 >100 comparative polysorbate 20 62 74 24 11 comparative polysorbate 80 60 74 nt 12 comparative n-dodecyl 62 nt nt 10 b-maltoside inventive Example 3 70 76 3.9.sup.(2) nt inventive Example 5 59 nt 18.7 nt inventive Example 2 72 68 9.9 nt inventive Example 4 75 84 0.5.sup.(2) 6.5 inventive Example 1 64 73 10.8 nt note.sup.(2): values of 4 or less are considered to be equivalent to zero in view of experimental variability.

(7) In the T.sub.onset results, all the inventive examples showed comparable or superior stability to the comparative examples, and Examples 2, 3, and 4 were especially superior to the comparative examples. In the Tm results, all the inventive examples showed comparable or superior stability to the comparative examples, and Example 4 was especially superior to the comparative examples. In the CD results, all the inventive examples showed results far superior to the comparative samples. In the D2 k results, the only inventive example tested, Example 4, was superior to the comparative examples.

EXAMPLE 2

Testing of Protein Solutions Containing IgG

(8) Scanning DLS was performed on protein solutions containing 1 mg/mL IgG from rabbit, at 0.05 C./min from 25 C. to 80 C., with surfactants all 1 mg/mL. T.sub.onset was 66 C. for all samples, but, as temperature was increased above T.sub.onset, the sample using Example 1 showed much slower growth of particle diameter than all the other samples.

(9) Isothermal DLS was performed at 1 mg/mL rabbit IgG at 65 C., and size at 4000 seconds (D4 k) was reported. CD technique using human IgG compared change in absorbance at 218 nm due to heat aging at 65 C. for 18 hours. Results were as follows:

(10) TABLE-US-00003 Type Surfactant D4k (nm) CD (mdeg) comparative none >200 22 comparative polysorbate 20 22 21 comparative polysorbate 80 30 nt comparative alkyl maltoside 30 nt inventive Example 3 12.2 15.5 inventive Example 5 40 nt inventive Example 2 30 16 inventive Example 4 25 18 inventive Example 1 25 17.5

(11) In the CD results, the inventive examples were superior to the comparative examples. In the D4 k results, Example 3 was superior to the comparative examples.

EXAMPLE 3

Results using Orencia Powder

(12) ORENCIA (abatacept) powder (manufactured by Bristol-Myers Squib) was obtained by prescription. Orencia was diluted with water to 4 mg/mL and tested by scanning DLS for T.sub.onset. Also, Orencia was diluted to 10 mg/mL with water and tested by isothermal DLS at 73 C., and the diameter was noted at 20 hours (D20 h). In protein solutions, D20 h tends to grow larger as time passes, and less-stable solutions reach higher values of D20 h. Smaller value of D20 h means a more stable protein solution. Orencia solutions at 1 mg/mL were tested in an isothermal step experiment; particle diameter was measured by DLS as a function of time during a hold of 45 hours at 50 C., followed by 15 hours at 60 C. The diameter at the end of this process, Dstep, was recorded. Smaller values of Dstep means a more stable protein solution.

(13) Results were as follows.

(14) TABLE-US-00004 Type Surfactant T.sub.onset ( C.) D20 h (nm) Dstep (nm) comparative none 62 >100 >100 comparative polysorbate 20 62 6 5 comparative polysorbate 80 53 7 5.5 comparative poloxamer 188 70 6 5.1 comparative alkyl maltoside 69 4.9 4.1 inventive Example 3 >80 5.6 4.7 inventive Example 5 72 5.7 5.1 inventive Example 4 75 5.4 4.9

(15) In the T.sub.onset and D20 h results, the inventive samples had more stable protein solutions than the comparative samples.

EXAMPLE 4

Test Results on Remicade

(16) REMICADE (infliximab) powder (manufactured by Johnson and Johnson) was obtained by prescription. Remicade powder was diluted to 1 mg/mL in water. Using scanning DLS, T.sub.onset was measured as described above. Also, isothermal DLS was performed at 50 C., and the time at which the particle diameter began to grow rapidly (t-grow) was noted. Results were as follows:

(17) TABLE-US-00005 Type Surfactant T.sub.onset ( C.) t-grow (min) comparative none 54 <10 comparative polysorbate 20 53 1400 comparative polysorbate 80 54 1400 comparative alkyl maltoside 53 625 inventive Example 3 53 535 inventive Example 5 53 1100 inventive Example 2 53 250 inventive Example 4 53 175 inventive Example 1 53 870

(18) The inventive samples showed comparable performance to the comparative samples.