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2004 American Diabetes Association 64th
Scientific Sessions |
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Alveair™, A New Pulmonary Insulin - Its Glucose
Responses Compared to Aspar Insulin Intravenous Injection in Normal
Rats and Its Insulin Profile in Guinea Pigs
Jing Li, M.D. , Frank K Leung, M.D. , Sieting Wong, Ph.D. , Emily
Leung, B.A. and Shao L Su, M.D. Lake Bluff, Illinois, United States
.
This study is to evaluate the glucose responses of Alveair
insulin comparing to Aspar intravenous injection (i.v.) in normal
rats with 0.5 u/kg body weight dose and the insulin profile in
guinea pigs with 1 unit/kg. Procedures and materials: Alveair
insulin concentration was 2.7 u/ml as determined by
radioimmunoassays. Alveair insulin was pulmonary-delivered via
intra-trachea through a surgical incision. Aspar was injected into
the Jugular vein. The Aspar group had otherwise identical
procedures. The Aspar i.v. identical dosing volume and insulin
concentration as the Alveair group. 4 normal rats were used for each
group.Tail blood glucose was sampled at time 0, 15, 30, 60, 120,
180, 240, 300, 360, 420 and 480 minutes.In a different experiment
and identical procedures, insulin levels were sampled from the
Jugular vein at time 0, 5, 15,30,60,120, 180 minutes in guinea pigs.
Statistics: SAS 8.01. Significance at 5% confidence. Results: %
glucose level compared to initial fasting glucose: Alveair - 100,
97, 89, 78,32,25,29,30,35,39. Aspar i.v. -
100,91,72,57,36,32,31,21,26,25,28.The maximal glucose reduction
compared to initial fasting glucose: Alveair - 75%; Aspar - 79%. The
nadir occurred between 120-180 minutes for both Alveair and Aspar
i.v. groups. Aspar i.v. had quicker onset of action than Alevair
group.Insulin samplings in guinea pigs (Microunit/ML): Asp -
0,15,311,374,110,16,0. Alveair - 0,20,45,100,165,275,320.
Conclusion:Aspar i.v. has quicker onset of action. In evaluating %
of glucose reduction compared to initial fasting values and the
duration of action, Alveair is similar and comparable to Aspar
intravenous injection in its efficacy. The time kinetics of insulin
release is quite different in Alveair than Asp as studied in guinea
pig. Alveair has a prolonged and slow release of insulin over time.
Its insulin bioavailability is similar to Asp intravenous injection
in this study. However, additional experiments will be performed to
confirm this interesting finding and reported later on in the
future. |
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Insulin Pharmacokinetic Estimates of Alveair™ - A New
Pulmonary Insulin Formulation
Frank K Leung, M.D. *, Sieting Wong, Ph.D. , Jing Li, M.D. ,
Emily Leung, B.A. and Norman Liu, Ph.D. . Lake bluff, Illinois .
The study is to evaluate the Alveair™ insulin PK estimates in
comparison to Aspar subcutaneous injection (Asp) in normal rats,
tested with two different dosages and insulin concentrations: 1) 2
units/kg body weight - Alveair insulin concentration was 2.7
units/ml as determined by radioimmunoassays. Aspar was 100 units/ml.
2) 0.5 unit/kg body weight - Alveair was 2.7 units/ml. Aspar was
diluted with phosphate buffer to 2.7 units/ml such that both drugs
were given with the same dosing volume and concentration. Procedure:
Concurrent study was performed on normal rats (average weight: 350
gm, n=2 for each dose) after 14 hours fast. All insulin measurements
were performed at Quest Diagnostic Laboratory and Coremeds
laboratory. Alveair insulin was pulmonary-delivered via
intra-trachea through a surgical incision. Insulin was sampled at
time 0, 15, 30, 60, 120 and 180 minutes. Statistics: SAS 8.01
system. Results: Dose, Tmax, Cmax, AUC_T, AUC_average. 1) 2 u/kg
Dose - Alveair: 60, 492, 30517, 169.54. Aspar: 60,197,14227, 79.042.
2) 0.5 u/kg Dose - Alveair: 15, 46.7, 2301, 12.784. Aspar: 15, 46.7,
412.5, 13.75. Summary: 1) Alveair insulin bioavailability based on
AUC_ave was 214% of Aspar sc at 2 u/kg dose. When Aspar was diluted
to the same insulin concentration and dosing volume as Alveair,
Alveair insulin bioavailability was 93% of Aspar sc. 2) Similarly,
Alveair Cmax is equal or better than Aspa. 3) For both formulations,
Tmax was 15 minutes at 0.5 u/kg dose and 60 minutes at 2 units/kg.Conclusion:
The insulin PK estimates may vary depending on the dosing volume and
insulin concentration of the formulations. In this study, Alveair
insulin bioavailability is close to 100% or better than Aspar
subcutaneous injection in normal rat |
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Dose-Responseness of Alveair™ Insulin in Normal Rats
- A New Pulmonary Insulin Formulation.
Frank K Leung, M.D. *, Sieting Wong, Ph.D. , Jing Li, M.D. and
Emily Leung, B.A.. Lake Bluff, Illinois, United States .
Purpose: To determine the dose-responsiveness of Alveair insulin
in normal rats. Procedures:Two concentrations were used in this
study: 0.22 unit/ml was used for 0.1 and 0.2 unit/kg body weight
dosages. 2.7 units/ml was used for all other dosages. Alveair
insulin was pulmonary-delivered via intra-trachea through a surgical
incision.Dosages tested: 0.1, 0.2, 0.5, 1 and 3 units/kg body
weight. 1 unit/kg dosage group had 8 rats, all others had 4 rats in
each group. Rats weights were 175 to 300 grams, matched with
placebo. All rats were fasted 14 hours overnight with normal fasting
glucose levels less than 130 mg/dl. Statistics: SAS 8.01.
Significance at 5% level. Results: Dosage, Slope (s.e.), AUC_average,
AUC_t- 1) 0.1 u/kg,-0.3(0.06), 62.55, 34025. 2) 0.2
u/kg,-0.31(0.06), 62.77, 30132, 3) 0.5 u/kg,-0.37(0.06), 52.31,
25110, 4) 1 u/kg,-0.45(0.06), 46.69, 22413, 5) 3 u/k,-0.41(0.064)
26.10, 12530. 6) Placebo,-0.15(0.064), 84.78, 31094, 7) Aspar (sc)
0.5 u/kg,-0.27(0.055), 64.78, 31094, 8) Aspar sc, 1
u/kg,-0.29(0.055), 76.26, 36606, 9) Aspar sc, 2 u/kg,-0.41(0.064),
48.39, 23228. Significant hypoglycemic action occurred at 60 minutes
in all Alveair groups with nadir at 2-3 hours. All slopes of Alveair
doses were significant different from placebo (P<0.0001).Percent
glucose reduction compared to initial fasting levels: 0.1 - 0.5 u/kg
by 30 to 60%; 0.5 - 1 u/kg by 50 to 70%; 1-3 u/kg by 60 to 90%. At
0.1 u/kg dose, the average glucose reduction was 40% from its
initial fasting level. Glucodynamic profiles were similar in all
dosage groups. The nadir occurred at 2nd to 3rd hour and was
sustained at steady levels through the study period by the Alveair
formulations. Aspar sc has quicker onset of action than Alveair. The
glucose levels of Aspar sc bounced back more than the Alveair
groups. Conclusion: Alveair insulin is a very efficacious pulmonary
formulation and significant glucose reduction occurred at
sub-unit/kg body weight dosages. It is at least as effective as
Aspar insulin subcutaneous injection in its efficacy. |
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Alveair™, A New Pulmonary Insulin - Its Glucodynamic
Profile Comparing to Aspar Insulin Subcutaneous Injection in Normal
Rats
Frank K Leung, M.D. *, Sieting Wong, Ph.D. , Jing Li, M.D. and Emily
Leung, B.A. Lake Bluff, Illioins, United States .
This study to evaluate the glucose responses of Alveair insulin
compared to Aspar Subcutaneous Injection (Asp) in normal rats with 2
insulin dosages (0.5 u/kg, 1 u/kg) and 2 Aspar insulin
concentrations (100 u/ml,2.7 u/ml). Procedures and materials:
Alveair insulin concentration was 2.7 u/ml as determined by
radioimmunoassays. Alveair was pulmonary-delivered via intra-trachea
through a surgical incision.Alveair at 1 u/kg had 8 rats. All other
groups had 4.Tail blood glucose was sampled at time 0, 15, 30, 60,
120, 180, 240, 300, 360, 420 and 480 minutes. Statistics: SAS 8.01.
Significance at 5% confidence. Results: 1) % glucose levels -
Alveair (0.5u/kg) - 100, 109, 113, 104, 79, 62, 50, 43, 42, 41, 44.
Aspar (0.5u/kg) :100,109,97,85,65,52,51,67,68,70,77,81. Alveair
(1u/kg):100,105,111,89,59,37,33,35,34,38,44,50.Aspar (1u/kg): 100,
102, 104, 92, 67, 50,55,65,104,102,103. 2) (Dose,Slope estimate(s.e.),
AUC_Averge, AUC_T). Alveair: (0.5u/kg,-0.37(0.055), 52.31, 25110);
(1u/kg, -0.31(0.055),46.69, 22413). Aspar sc: (0.5
u/kg,-0.27(0.055), 64.78, 31094); (1u/kg,-0.29(0.055), 76.26,36606).
Placebo: (0 u/kg,-0.15(0.06), 84.78, 40765). The nadir occurred
120-180 minutes for all groups.Slopes estimates were comparable
between Alveair and Asp. The glucose started to rebound
significantly towards initial glucose levels following the nadir in
Aspar groups and more so in the Aspar group receiving 100 u/ml
insulin concentration, but the glucose in the Alveair groups
remained low and started to rebound slightly at 420 to 480 minutes.
The Aspar group with 0.5 u/kg dose had a quicker onset of action
than Alevair groups. The maximal glucose reduction compared to the
initial fasting glucose in the Aspar groups averaged 50%; the
Alveair 0.5u/kg group averaged 59% and the Alveair 1 u/kg group
averaged 67%. Conclusion: In this study, Aspar at diluted insulin
concentration has quicker onset of action than Alveair. The Alveair
efficacy and AUC_glucose were better than the Aspar groups, also due
to the fact that the glucose in the latter groups tend to rebound
earlier. |
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Alveair™, A New Pulmonary Insulin - Its First Human
Trial and The Pre-clinical Animal Data Correlation
Frank K Leung, M.D. *, Martin C Fuh, M.D. *, Cheng C Lin, M.D. ,
Sieting Wong, Ph.D. , Jing Li, M.D. , Emily Leung, B.A. , Norman
Liu, Ph.D. and Lang Ren, B.Pharm. Lake Bluff, Illinois, United
States ; Tai Chung, Taiwan and Taipei, Taiwan .
The study is to determine Alveairs effective threshold dosage.
Pre-clinical data: Alveair insulin (2.7 u/ml) was
pulmonary-delivered via intrac-trachea. Each group of 4 rats
received one of the doses: 0.1, 0.2, 0.5, 3 u/kg. Another group of 8
rats received 1 u/kg.Tail blood was sampled for glucose at
0,15,30,60,120,180,240,300,360,420,480 minutes.Results (SAS
8.01):Dosage, AUC_average, AUC_t- 1) 0.1 u/kg, 62.55, 34025. 2) 0.2
u/kg, 62.77, 30132, 3) 0.5 u/kg, 52.31, 25110, 4) 1 u/kg, 46.69,
22413, 5) 3 u/k, 26.10, 12530. 6) Placebo, 84.78, 31094. % glucose
reduction compared to initial levels: 0.1 - 0.5 u/kg by 30 to 60%;
0.5 - 1 u/kg by 50 to 70%; 1-3 u/kg by 60 to 90%.The nadir occurred
at 2 -3 hour.Human trial protocol: Single dose, two periods
cross-over design with an in-between wash-out period. Organ systems
for symptoms was reviewed before, during and after the study. 4
fasted, non-obese and healthy subjects (2 women, 2 men, 2 smokers)
received 75 gm carbohydrate orally, immediately followed by
inhalation of Alveair at 0.15 unit/kg (16.7 u/ml concentration). The
hand-held device vaporized Alveair with the median mass median
diameters (MMD) at 1.9 micron.End-point of the study: 1) when
glucose drops 40% of the initial fasting level or 2) any significant
symptoms occur.Results: All subjects developed hypoglycemic symptoms
(3-6 on 1-10 scale) between 2-4 hours, and none with placebo. Low
blood glucose was immediately corrected and normalized. % glucose
reduction from fasting levels: Alveair :(-46.3, -51.2, -46, -29.1).
Placebo: (-19.9, -18.1, -21, -22.5) Mean % reduction: Alveair (-43),
Placebo (-20), the difference was statistically significant (P
<0.03, SAS 8.01). There was not a single incidence of induced
coughing, sneezing, sinus or pulmonary irritations and allergic
reactions. Conclusion: Alveair was well tolerated in its first human
trial. There was significant glucose-lowering action at sub-unit/kg
dosage. The results were comparable to its pre-clinical animal data. |
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Effects of Immediate Food Intake Following Gavage
Delivery of Intesulin (oral insulin) in Diabetic Rats
Frank K Leung, M.D. *, Sieting Wong, M.D., Jing Li, M.D. and Emily
Leung, B.A. Lake Bluff, Illinois, United States .
The study is to evaluate the effects of immediate food intake
following Intesulin delivery by gavage method. Each group has 4
diabetic rats induced by Streptozocin. Exp1) compare Placebo to
Intesulin, immediately followed by one ml of Boost (20% protein, 14%
carbohydrate, 6% fat) in diabetic rats with fasting glucose 300-400
mg/dl. Exp 2) as in experiment 1, except the diabetic rats had
fasting glucose 90-150 mg/dl.The weight and fasting glucose were
matched for both placebo and Intesulin groups. All rats were fasted
for 18 hours. The study was concurrent. All experimental rats were
given the same volume of 0.2 ml of Intesulin orally. Control rats
were given 0.2 ml of placebo. The Intesulin dosage was approximately
2 to 3 units/kg depending on the weight. Tail blood was sampled for
glucose at time 0,15,30,60,120,180,240,300,360,420,480 minutes.
Results 1) % glucose levels: Placebo (Exp.1) - 100,104,99,102,
101,98,97,93, 96,99,95. Intesulin (Exp. 1) -
100,96,91,82,75,67,62,60,58,63,61. Placebo (Exp.2) -100,121, 131,
153, 184,188,181,183,171,166,161. Intesulin (Exp.2) 100,114,118,111,
98,104,100, 103, 112,101,95. 2) (Slope estimate(s.e.),T max mean, C
max mean, AUC_mean) Exp. 1: Placebo (0,170, 105,94.297), Intesulin
(-0.17(0.04),30,90.75,65.461). Exp. 2: Placebo
(0,195,202.5,167.422).Intesulin
(-0.1(0.066),126,130,99.418).Summary: Ratio of AUC_mean (Intesulin)/
AUC_mean (Placebo): Exp 1 = 69.4%, Exp.2 = 59.3%. Statistically
significant (SAS 8.01 significance at 5% level) hypoglycemic level
occurred from 120 to 480 minutes for Intesulin (Exp.1) and 120 to
360 for Intesulin (Exp.2) . Slopes comparisons were significantly
different for Exp.1 (P<0.02) and Exp.2 (P<0.0001). Conclusion:
Intesulin is effective in lowering blood glucose in the presence of
food, as evaluated by gavage method. The onset of significant
hypoglycemia action began 2 hours after food intake and could last
for 5 to 7 hours. |
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Effects of Food Intake Thirty Minutes Following
Gavage Delivery of Intesulin (oral insulin) in Diabetic Rats
Frank K Leung, M.D.*, Sieting Wong, Ph.D. , Jing Li, M.D. and Emily
Leung, B.A. . Lake Bluff, Illinois, United States .
The study is to evaluate the effects of food intake 30 minutes
following Intesulin delivery by gavage method. Experiment and
procedures: Each group has 4 diabetic rats induced by
Streptozocin.One ml of Boost (20% protein, 14% carbohydrate, 6% fat)
was given 30 minutes after delivery of Intesulin in diabetic rats by
gavage.The study was concurrent. The weight and fasting glucose were
matched for both placebo and Intesulin groups. All rats were fasted
for 18 hours. Tail blood was sampled for glucose at time 0,15,30,
60,120, 180,240,300,360,420,480 minutes.All experimental rats were
given the same volume of 0.2 ml of Intesulin orally. The dosage of
Intesulin was approximately 2-3 units/kg depending on the weight.
Results (Slope estimate (s.e.),T max mean, C max mean, AUC_mean)
Placebo (0.30(0.07),140, 169,147.51), Intesulin (-0. 27(0.06), 150,
102. 25,70.186). Mean percent glucose levels: Placebo:100,111,133,
141,166,164, 158,162, 148,143,139. Intesulin:
100,101,90,84,69,55,75,81,86,85,88. Summary: Slope comparison to
placebo was significantly different (P<0.0001). There was a
reduction of 52% of the AUC_ mean glucose compared to placebo group.
The maximal glucose reduction compared to the initial fasting
glucose was 45% in the Intesulin group. Statistically significant (SAS
8.01 significance at 5% level) hypoglycemic levels occurred at 30 to
300 minutes comparing to placebo.Conclusion: Intesulin is effective
in lowering blood glucose 30 minutes preceding food intake, as
evaluated by gavage method. The onset of significant hypoglycemic
action began 30 minutes after food intake and lasted for 5 hours. |
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Effects of Immediate Food Intake Following
Orally-delivered Intesulin by Gavage method and Its Comparison to
Aspar Subcutaneous Injection in Diabetic Rats
Frank K Leung, M.D.*, Sieting Wong, Ph.D., Jing Li, M.D. and Emily
Leung, B.A. . Lake Bluff, Illinois, United States .
The study is to evaluate the effects of immediate food intake
following Intesulin oral delivery compared to Aspar subcutaneous
injection (Asp) by gavage method. Diabetic rats were induced by
Streptozocin. Exp1) compare Asp (n=3) to Intesulin (n=4),
immediately followed by one ml of Boost (20% protein, 14%
carbohydrate, 6% fat) in diabetic rats with fasting glucose of
300-400 mg/dl. Exp 2) as in experiment 1, except the diabetic rats
had fasting glucose 90-150 mg/dl.The weight and fasting glucose were
matched for both Asp and Intesulin groups. All rats were fasted for
18 hours.All experimental rats were given the same volume of 0.2 ml
of Intesulin orally. Asp rats received 2 units/kg dose
subcutaneously. Intesulin dosage was approximately 2 to 3 units/kg
depending on the weight. Tail blood was sampled for glucose at time
0,15,30,60,120,180,240,300,360,420,480 minutes. Results 1) % glucose
levels: Asp (Exp.1) - 100,81,60,41,20,16,23,37,39,40,36. Intesulin
(Exp. 1) - 100,96,91,82,75,67, 62,60,58,63,61. Asp (Exp.2)
-100,110,85,47,30,27,22,28,43,57,66. Intesulin (Exp.2)
100,114,118,111,98,104,100,103,112,101,95. 2. 2) (Sope estimate(s.e.),
T max mean, C max mean, AUC_mean) Exp. 1: Asp
(-0.35(0.076),30,64.8,25.047), Intesulin (-0.15 (0.064),
30,90.75,65.461). Exp. 2: Asp (-0.91(0.11), 95,86.5,42.634).
Intesulin (-0.52 (0.1), 126,130,99.418). Summary: 1) Ratio of
AUC_mean (Intesulin)/ AUC_mean (Asp): Exp 1 = 261%, Exp.2 = 233%. 2)
Statistically significant (SAS 8.01 significance at 5% level)
hypoglycemic level occurred from 120 to 480 minutes for Intesulin
(Exp.1) and 120 to 360 for Intesulin (Exp.2); 15 -480 minutes for
Asp (Exp.1) and 30 -480 minutes for Asp (Exp.2).3).Conclusion: Both
Intesulin and Asp are effective in lowering blood glucose in the
presence of food, as evaluated by gavage method. Aspar subcutaneous
injection has quicker onset of hypoglycemic action and is
approximately 2-3 times more efficacious than orally-delivered
Intesulin in this study. |
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Glucose Responses to Oral Delivery of Intesulin™ by
Gavage Method in Guinea Pigs
Jing Li, M.D. , Frank K Leung, M.D. *, Sieting Wong, Ph.D., Emily
Leung, B,A. and Shao L Su, M.D. . Lake Bluff, Illinois, United
States .
We have previously demonstrated the hypoglycemic efficacy of
Intesulin oral insulin in rats using gavage method to deliver the
formulation directly into the gastric cavity.This study is to
evaluate the glucose responses of Intesulin oral insulin in
different animal species using guinea pig as a model. Procedures and
materials: Intesulin insulin concentration was 2.7 u/ml as
determined by radioimmunoassays at Quest Diagnostic Laboratory and
Coremeds laboratory. 5 units/kg body weight dose was delivered into
the stomach via gavage needle (Harvard Apparatus). Controls received
similar dose volume of placebo. 4 normal and non-diabetic guinea
pigs were used for each group. All animals were fasted overnight for
18 hours. Blood glucose was sampled at time 0, 15, 30, 60, 120, 180,
240, 300, 360, 420 and 480 minutes from the ears. Statistics: SAS
8.01. Significance at 5% confidence. Results: % glucose levels
compared to initial fasting glucose: Intesulin -
100,90,74,58,37,28,29,26,21,23,22.. Placebo -
100,98,104,99,94,83,63,59,54,48,43. Standard errors were comparable
between the two groups. Significant glucose-lowering action occurred
at 30 minutes. Glucose reduction compared to initial fasting glucose
- At 120 minutes: Intesulin (- 63%); Placebo (-27%). At 300 minutes:
Intesulin (-74%), Placebo (-41%). At 360-480 minutes, glucose in the
placebo group dropped significantly due to prolonged starvation. But
the glucose in the Intesulin was still significantly lower than the
placebo. The nadir occurred between 120-180 minutes for Intesulin
group. Conclusion: At 5 units/kg dose, Intesulin is very effective
in lowering the glucose levels of fasting non-diabetic guinea pigs,
delivered directly into the gastric cavity via gavage method. Its
hypoglycemic effect can last for many hours. |
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