Journal: Pharmacy and Therapeutics

Article Title: Pharmacological Agents Utilized in Patients With Type-2 Diabetes: Beyond Lowering A1c

doi:

Figure Lengend Snippet: Summary of FDA-Approved Medications for Use in Patients With Type-2 Diabetes 31

Article Snippet: Pramlintide , SymlinPen (AstraZeneca) , SQ pen , 1,000 mcg/mL, 1.5–2.7 mL (2s) , $235.81– $357.44.

Techniques:

Journal: bioRxiv

Article Title: Ultra-fast insulin-pramlintide co-formulation for improved glucose management in diabetic rats

doi: 10.1101/2021.04.12.439573

Figure Lengend Snippet: a, Current dual-hormone replacement of insulin and pramlintide requires two separate injections at mealtimes (idealized data for illustration based on reported pharmacokinetics). Not only is this additional injection burdensome, but there is a kinetic mismatch between insulin and pramlintide when delivered exogenously compared to endogenous co-secretion from the beta-cells. This results from the mixed insulin association states present in rapid-acting insulin formulations where monomers and dimers are rapidly absorbed, but the slow dissociation of the insulin hexamer causes extended duration of action. b, A single injection co-formulation of monomeric insulin and pramlintide would reduce patient burden, and have better pharmacokinetic overlap that more closely mimics endogenous secretion from the healthy pancreas (idealized data for illustration of study goals). c, Amphiphilic acrylamide copolymer excipients can be used to stabilize an insulin-pramlintide co-formulation. These excipients preferentially adsorb onto the air-water interface, displacing insulin and/or pramlintide and preventing the nucleation of aggregation events that initiate amyloid fibril formation. d, Co-formulation components. e, Insulin association states in (i) Humalog (adapted from the literature 4 ) compared to (ii) zinc-free lispro with phenoxyethanol (0.85 wt.%) and glycerol (2.6 wt.%). f, Formulation stability in a stressed aging assay (continuous agitation, 37 °C) of (i) Humalog, (ii) Humalog + pramlintide (1:6 pramlintide:lispro), (iii) zinc-free lispro (100U/mL lispro, 0.85 wt.% phenoxyethanol, 2.6 wt.% glycerol, 0.1 mg/mL MoNi 23% ), (iv) Co-formulation (100 U/mL lispro, 1:6 pramlintide:lispro, 0.85 wt.% phenoxyethanol, 2.6 wt.% glycerol, 0.1 mg/mL MoNi23%). Change in transmittance is shown from baseline transmittance. Aggregation is defined as a change in transmittance >10%.

Article Snippet: [ ] Characterization of MoNi 23% molecular weight and monomer composition can be found in Table S1. in Humalog (Eli Lilly) and pramlintide (BioTang) were purchased and used as received.

Techniques: Injection

Journal: bioRxiv

Article Title: Ultra-fast insulin-pramlintide co-formulation for improved glucose management in diabetic rats

doi: 10.1101/2021.04.12.439573

Figure Lengend Snippet: Fasted male diabetic rats (n=11) received subcutaneous administration of (i) Humalog, (ii) separate injections of Humalog and pramlintide, or (iii) insulin-pramlintide co-formulation. a, Insulin administration was immediately followed with oral gavage with a glucose solution (1 g/kg). Each rat received all treatment groups. b, Change in blood glucose levels from baseline following treatment. c,d, Pharmacokinetics of ( c) insulin lispro or (d) pramlintide. See Figure S3 and S4 for area under the curve (AUC) exposure comparison for lispro (F 2,20 =0.53, P=0.59) and pramlintide (F 2,10 =3.27, P=0.10).

Article Snippet: [ ] Characterization of MoNi 23% molecular weight and monomer composition can be found in Table S1. in Humalog (Eli Lilly) and pramlintide (BioTang) were purchased and used as received.

Techniques:

Journal: bioRxiv

Article Title: Ultra-fast insulin-pramlintide co-formulation for improved glucose management in diabetic rats

doi: 10.1101/2021.04.12.439573

Figure Lengend Snippet: Fasted male diabetic rats (n=11) received subcutaneous administration of (i) Humalog, (ii) separate injections of Humalog and pramlintide, or (iii) insulin-pramlintide co-formulation. Insulin administration was immediately followed with oral gavage with a glucose solution (1 g/kg). Each rat received all treatment groups. a,j, Pharmacokinetics for each rat was individually normalized to the peak serum levels and the normalized values were averaged for (a) insulin lispro or (j) pramlintide. b,k, Exposure onset defined as time to 50% of the peak up for (b) insulin lispro or (k) pramlintide. c,l, Exposure peak for (c) insulin lispro or (l) pramlintide. d,m, Exposure onset defined as time to 50% of the peak up for (d) insulin lispro or (m) pramlintide. e-i, Fraction of lispro exposure as a ratio of AUC t /AUC 120 at e, t=6; f, t=15; g, t=30; h, t=45; i, t=60. Statistical significance was determined by restricted maximum likelihood repeated measures mixed model. Tukey HSD post-hoc tests were applied to account for multiple comparisons (b-i, k-m). Bonferroni post hoc tests were performed to account for comparisons of multiple individual exposure time points, and significance and a were adjusted (α= 0.01) (e-i).

Article Snippet: [ ] Characterization of MoNi 23% molecular weight and monomer composition can be found in Table S1. in Humalog (Eli Lilly) and pramlintide (BioTang) were purchased and used as received.

Techniques:

Journal: bioRxiv

Article Title: Ultra-fast insulin-pramlintide co-formulation for improved glucose management in diabetic rats

doi: 10.1101/2021.04.12.439573

Figure Lengend Snippet: a,b, Average normalized serum concentrations (for each rat, n=11/group) for insulin and pramlintide when delivered (a) as two separate injections and (b) when delivered together as a co-formulation. c, Overlap between the two curves was defined as the total time spent above 0.5 for both insulin and pramlintide curves (width at half-peak height), shown as a ratio of the overlap time to the total width of both peaks (overlap ÷ (lispro + pramlintide − overlap). Statistical significance was determined by restricted maximum likelihood repeated measures mixed model.

Article Snippet: [ ] Characterization of MoNi 23% molecular weight and monomer composition can be found in Table S1. in Humalog (Eli Lilly) and pramlintide (BioTang) were purchased and used as received.

Techniques:

Journal: bioRxiv

Article Title: Ultra-fast insulin-pramlintide co-formulation for improved glucose management in diabetic rats

doi: 10.1101/2021.04.12.439573

Figure Lengend Snippet: Fasted male diabetic rats received subcutaneous administration of (i) Humalog, (ii) separate injections of Humalog and pramlintide, or (iii) insulin-pramlintide co-formulation. a, Gastric emptying experiment where insulin administration (2 U/kg) was immediately followed with oral gavage with an acetaminophen slurry (100 mg/kg). Each rat (n=11) received all treatment groups. b, Acetaminophen serum concentration. c, Time to peak exposure of acetaminophen serum concentration. All data is shown as mean ± SE. Statistical significance was determined by restricted maximum likelihood repeated measures mixed model. Tukey HSD post-hoc tests were applied to account for multiple comparisons.

Article Snippet: [ ] Characterization of MoNi 23% molecular weight and monomer composition can be found in Table S1. in Humalog (Eli Lilly) and pramlintide (BioTang) were purchased and used as received.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: Ultra-fast insulin-pramlintide co-formulation for improved glucose management in diabetic rats

doi: 10.1101/2021.04.12.439573

Figure Lengend Snippet: Fasted male diabetic rats received subcutaneous administration of (i) Humalog, (ii) separate injections of Humalog and pramlintide, or (iii) insulin-pramlintide co-formulation. a, Oral glucose challenge where insulin administration (0.75 U/kg) was immediately followed with oral gavage with a glucose solution (2 g/kg). Each rat (n=10) received all treatment groups. b, Change in blood glucose after administration is shown. c, Max change in glucose above baseline d, Max change in glucose below the baseline. All data is shown as mean ± SE. Statistical significance was determined by restricted maximum likelihood repeated measures mixed model. Tukey HSD post-hoc tests were applied to account for multiple comparisons.

Article Snippet: [ ] Characterization of MoNi 23% molecular weight and monomer composition can be found in Table S1. in Humalog (Eli Lilly) and pramlintide (BioTang) were purchased and used as received.

Techniques:

Journal: The Journal of Clinical Investigation

Article Title: Human IAPP is a contributor to painful diabetic peripheral neuropathy

doi: 10.1172/JCI156993

Figure Lengend Snippet: ( A and B ) Fluorescence (indicates amount of amyloid fibrils) of thioflavin T ( A ) and transmission electron microscopy imaging (scale bars: 0.2 nm) ( B ) after 24 hours of incubation of hIAPP, mIAPP, or pramlintide. ( C ) Sensory neurons were treated with 100 nM hIAPP, mIAPP, pramlintide, or saline for 24 hours. The average neurite length per neuron was assessed and expressed as the percentage length per neuron of vehicle-treated neurons ( n = 8; n represents a DRG culture of 1 mouse). ( D ) Mitochondrial ROS level in cultured DRG neurons incubated with hIAPP or pramlintide (10, 100, and 1,000 nM). Measurements are per cell from 3 different cultures, n = 551–654 neurons per group. ( E and F ) Course of mechanical sensitivity ( n = 5) ( E ) and IENF density ( n = 7) on day 6 ( F ) after intraplantar injection of 1,000 fg hIAPP, mIAPP, and pramlintide in male and female WT mice. ( G ) Density of IENFs in skin of T2DM subjects ( n = 6) and non-T2DM controls ( n = 9). ( H ) Representative images of G stained for the pan-neuronal marker PGP9.5 and collagen IV (lines indicate the border between dermis and epidermis; white arrows represent the IENF; scale bar: 20 μm). ( I ) IAPP-positive oligomers in skin of T2DM subjects ( n = 6) and non-T2DM controls ( n = 9). ( J ) Representative images of I stained for collagen IV (C IV), IAPP, and oligomers (I11). IAPP- and oligomer-positive spots are indicated by arrowheads. Lines indicate the border between dermis and epidermis. ( C and D ) One-way ANOVA with Dunnett’s test; * P < 0.05, ** P < 0.01, *** P < 0.001. ( E and F ) Two-way ANOVA with Tukey’s test; * P < 0.05 vs. vehicle; ## P < 0.01 vs. pramlintide. ( G and I ) Unpaired t test; * P < 0.05, ** P < 0.01. Data are expressed as mean ± SEM.

Article Snippet: hIAPP, mIAPP (both obtained from Bachem), and pramlintide (obtained from AnaSpec) were dissolved in dH 2 O to obtain stock solutions of 200 μg/mL; aliquots were stored at –80°C.

Techniques: Fluorescence, Transmission Assay, Electron Microscopy, Imaging, Incubation, Saline, Cell Culture, Injection, Staining, Marker