Journal: Nature biomedical engineering

Article Title: A co-formulation of supramolecularly stabilized insulin and pramlintide enhances meal-time glucagon suppression in diabetic pigs

doi: 10.1038/s41551-020-0555-4

Figure Lengend Snippet: a, In vitro stability of pramlintide formulations at various pH values with and without CB[7]-PEG. b, In vitro stability of pramlintide-aspart (1:6 and 1:20 molar ratio) co-formulations with CB[7]-PEG at physiological pH. c, In vitro stability of pramlintide-lispro (1:6 and 1:20 molar ratio) co-formulations with CB[7]-PEG at physiological pH. Co-formulations were compared to controls of commercial Novolog or Humalog, and mixtures of the incompatible aspart+pramlintide or lispro+pramlintide in the absence of CB[7]-PEG. These assays assess the aggregation of proteins in formulation over time during stressed aging (i.e., continuous agitation at 37ºC) by monitoring changes in transmittance at 540nm. These experiments demonstrate that formulation with CB[7]-PEG prevents protein aggregation over the 100h period assayed, even when commercial formulations aggregate within 10h. Data shown are average transmittance traces for n = 3 samples per group.

Article Snippet: Simultaneously with their morning meal (200 g Teklad Miniswine Diet 8753; 66 g carbohydrates), pigs were injected subcutaneously with a 4U dose (0.13 U/kg) of the following formulations: (i) no treatment (pigs received food only) (ii) Humalog (100U/mL, Eli Lilly), (iii) separate administrations of Humalog and pramlintide (pH=4) (1:6 pramlintide to lispro, 0.5 μg/kg), (iv) lispro-pramlintide co-formulation (zinc-free lispro at 0.13 U/kg; pramlintide at 0.5 μg/kg) with CB[7]-PEG (3 eq to insulin + pramlintide).

Techniques: In Vitro, Formulation

Journal: Nature biomedical engineering

Article Title: A co-formulation of supramolecularly stabilized insulin and pramlintide enhances meal-time glucagon suppression in diabetic pigs

doi: 10.1038/s41551-020-0555-4

Figure Lengend Snippet: Diabetic female pigs received subcutaneous administration of therapies comprising either (i) commercial Humalog, (ii) commercial Humalog and pramlintide (pH=4) delivered in separate injections, or (iii) lispro-pramlintide co-formulation with CB[7]-PEG. Treatments were administered simultaneously with a 200g meal. All treatment groups received 4U insulin and pramlintide groups received a molar ratio of 1:6 pramlintide to lispro. Pharmacokinetics of a, insulin lispro in mU/L lispro (Humalog n=14; Separate n=15; Co-formulation n=13) or c, pramlintide in pM (Separate n=14; Co-formulation n=14). The area under the pharmacokinetic curves (AUC) of b, lispro (Humalog n=12; Separate n=14; Co-formulation n=13) and d, pramlintide (Separate n=13; Co-formulation n=14) for the first 240 minutes after subcutaneous injection. Pharmacokinetics for each pig were individually normalized to peak concentrations and normalized values were averaged for e, lispro (Humalog n=14; Separate n=15; Co-formulation n=13) or i, pramlintide (Separate n=14; Co-formulation n=14) concentration for each treatment group. Time to reach 50% of peak f, lispro (Humalog n=13;Separate n=14; Co-formulation n=12) or j, pramlintide concentration (onset) (Separate n=13; Co-formulation n=13). Time to reach peak g, lispro (Humalog n=14; Separate n=14; Co-formulation n=12) or k, pramlintide concentration (Separate n=13; Co-formulation n=14). Time for h, lispro (Humalog n=14; Separate n=15; Co-formulation n=13) or l, pramlintide depletion to 50% of peak concentration (Separate n=13; Co-formulation n=13). The specified sample size n refers to a cohort of 11 pigs who received each treatment group an equal number of times. Error bars, mean ± s.d. The Grubbs’ outlier test (alpha=0.05) was used to remove outliers. Statistical significance was determined by a two-tailed student’s t-test.

Article Snippet: Simultaneously with their morning meal (200 g Teklad Miniswine Diet 8753; 66 g carbohydrates), pigs were injected subcutaneously with a 4U dose (0.13 U/kg) of the following formulations: (i) no treatment (pigs received food only) (ii) Humalog (100U/mL, Eli Lilly), (iii) separate administrations of Humalog and pramlintide (pH=4) (1:6 pramlintide to lispro, 0.5 μg/kg), (iv) lispro-pramlintide co-formulation (zinc-free lispro at 0.13 U/kg; pramlintide at 0.5 μg/kg) with CB[7]-PEG (3 eq to insulin + pramlintide).

Techniques: Formulation, Drug discovery, Injection, Concentration Assay, Two Tailed Test

Journal: Nature biomedical engineering

Article Title: A co-formulation of supramolecularly stabilized insulin and pramlintide enhances meal-time glucagon suppression in diabetic pigs

doi: 10.1038/s41551-020-0555-4

Figure Lengend Snippet: Pharmacokinetics of lispro and pramlintide after injection with Humalog and pramlintide as separate injections and as a lispro-pramlintide co-formulation. Mean normalized concentration (normalized individually for each pig) of lispro and pramlintide when administered as a, two separate injections (Humalog n=15; Pramlintide n=13) or b, as a co-formulation (Lispro n=13; Pramlintide n=14) with CB[7]-PEG. The overlap between curves was evaluated as the time during which both lispro and pramlintide concentrations were greater than 0.5 (width at half peak height), shown as a ratio of c, overlap time over the total width of both peaks (Overlap/(Lispro + Pramlintide - Overlap)) (Separate n=13; Co-formulation n=13). Pharmacokinetics of glucagon after a meal and treatment with Humalog alone, Humalog and pramlintide as separate injections or as a lispro-pramlintide co-formulation. Glucagon is plotted as d, change in glucagon concentrations from baseline over 4-hours following treatment administration e, overall distance from baseline by treatment group (sum of individual points) (Humalog n=13; Separate n=14; Co-formulation n=14). The co-formulation reduced glucagon levels compared to Humalog (*p=0.0465) and separate administrations of Humalog and pramlintide (*p=0.0495). f, A summary schematic of how treatment affects post-prandial glucagon. The specified sample size n refers to a cohort of 11 pigs who received each treatment group an equal number of times. Error bars indicate mean ± s.d. The ROUT test (Q=1%) was used to remove outliers. Statistical significance was determined by a two-tailed student’s t-test.

Article Snippet: Simultaneously with their morning meal (200 g Teklad Miniswine Diet 8753; 66 g carbohydrates), pigs were injected subcutaneously with a 4U dose (0.13 U/kg) of the following formulations: (i) no treatment (pigs received food only) (ii) Humalog (100U/mL, Eli Lilly), (iii) separate administrations of Humalog and pramlintide (pH=4) (1:6 pramlintide to lispro, 0.5 μg/kg), (iv) lispro-pramlintide co-formulation (zinc-free lispro at 0.13 U/kg; pramlintide at 0.5 μg/kg) with CB[7]-PEG (3 eq to insulin + pramlintide).

Techniques: Drug discovery, Injection, Formulation, Concentration Assay, Two Tailed Test