Method for nasal application of a medicinal substance

Abstract

A method for nasal application of a medicinal substance by applying the substance through the nose in a maximum amount that is insufficient to stimulate an excretory response that would clear a significant portion of the substance from nasal and sinus passages. Within a time period of less than one hour, the application of the substance through the nose in an amount that is insufficient to stimulate an excretory response that would clear a significant portion of the substance from nasal and sinus passages is repeated. The repeated application, at a minimum, is done a sufficient number of times to provide an effective total dose of the substance. The repeated application, in any case, is done at least once.

Description

BACKGROUND OF THE INVENTION

[0002] This invention relates to delivery of a medical substance to a mammal and more particularly relates to nasal delivery of such a substance for absorption, reaction or other utilization.

[0003] Nasal administration of medical substances has had significant disadvantages. Dosages have been difficult to control due to excretory responses to administration, e.g. sneezing and mucosal excretion that removes or significantly reduces the substance from the nasal passages and sinuses. Another problem is that administered substances removed by an excretory response are often swallowed leading to nausea, stomach upset or other digestive disturbance. An even more serious problem is that if the excretory response is strong enough, the substance can be inhaled causing coughing or more serious pulmonary distress. An even further problem is that when a significant amount of the medical substance is removed by an excretory response, the material excreted is wasted material, thus increasing costs and inefficiencies associated with nasal administration.

BRIEF DESCRIPTION OF THE INVENTION

[0004] In accordance with the invention a method is therefore provided for nasal application of a medicinal substance which overcomes the above disadvantages. In particular, the method comprises applying the substance through the nose in a maximum amount that is insufficient to stimulate an excretory response that would clear a significant portion of the substance from nasal and sinus passages and within a time period of less than one hour, and repeating the application of the substance, through the nose in an amount that is insufficient to stimulate an excretory response that would clear a significant portion of the substance from nasal and sinus passages. The repeated application, at a minimum, is done a sufficient number of times to provide an effective total dose of the substance. The repeated application, in any case, is done at least once.

DETAILED DESCRIPTION OF THE INVENTION

[0005] “Nasal application”, as used herein, means applied through the nose into the nasal or sinus passages or both. The application may, for example, be done by drops, sprays, mists, coatings or mixtures thereof applied to the nasal and sinus passages.

[0006] “Medicinal substance” means any substance capable of being effectively applied nasally. Such substance are usually in the form of liquids, but may also be vapors or fine solids. Such substances are either absorbed by the tissues and vessels in the nasal and sinus passages (nasally absorbable) or interact with the surface of such passages (nasally active). Such substances may for example include vaccines, antigens, epitopes, adjuvants, viral vectors, bacterial vectors, immune modulators, delivery vehicles, and other drugs such as antibiotics, antivirals, hormones, antibodies, anti-inflammatories, antipyretics, antispasmotics, sedatives, anesthetics, chemotherapeutic agents, analgesics, vasodialators, and vasoconstrictors.

[0007] When the medical substance is a vaccine it may for example be a vaccine for non-typeable haemophilus influenzae which may contain an epitope of P5, P6 or both P5 and P6 proteins of haemophilus influenzae. The vaccine may also for example be a vaccine against hepatitis B.

[0008] The maximum amount that is insufficient to stimulate an excretory response that would clear a significant portion of the medicinal substance from the nasal and sinus passages is readily determined by observation and varies with the substance being applied, the surface area of the nasal passages and sinuses and with the size and species of animal. In the case of a mouse, the maximum amount is usually between about 2 and 10 μl and for a human is usually from about one to about three drops.

[0009] “Excretory response” means a response by the animal that tends to clear a significant portion of the medicinal substance from the nasal passages and sinuses. Such responses include increased secretions from the surfaces of the nasal passages and sinuses, and sneezing. Increased secretions may dilute the substance and can be removed from the nasal passages and sinuses by sneezing, blowing, dripping, coughing and swallowing.

[0010] “Significant portion” means that the effectiveness of the substance is substantially reduced (e.g. a reduction in effectiveness greater than 20 percent) due to excretion. A “significant portion” would normally be between 10 and 30 percent of the applied dose.

[0011] Repeated applications to obtain a maximum dose without stimulating an excretory response, for practical reasons related to the value of doctor and patient time, are usually completed within an hour and preferably less, e.g. one-half hour. The total number of doses within an hour is at least two but to obtain maximum effective dose, usually the number of doses is between 3 and about 20 and preferably between 4 and about 12 within an hour. Commonly, the number of applications is from 3 to about 15 applications within an hour. The time interval between doses is usually between about 30 seconds and about 15 minutes.

[0012] The method of the invention is applicable to essentially any mammal having easily accessible nasal passages and sinuses, e.g. mice, rats, chinchillas and other rodents, cats, monkeys, apes and humans. It has been found that position of certain mammals may increase effectiveness. For example, application is more effective in a prone chinchilla than a supine chinchilla and more effective in a supine mouse than a prone mouse. Nevertheless, the method of the invention using repeated doses, below the amount that stimulates a significant excretory response, is more effective than single doses when other variables are constant.

[0013] The following examples serve to illustrate but not limit the invention.

[0014] To show the distribution of liquid administered through the nose, Evans Blue Dye (0.3%) was administered through a micropipette tip into the nose of mice and chinchillas at various doses, at various levels of sedation or anesthesia, and with the animals in various positions.

[0015] The results clearly show that when a lower dose is used, more dye is retained in the nasal passages and sinuses and less dye is lost to the esophagus, stomach, intestines and lungs. The results further clearly show that when a series of low doses are used near the point at which the animal excretes the dye to the esophagus, stomach and intestines, more material can be retained in the nasal passages and sinus cavities than when a single larger dose is used. Further interesting results are that more dye is retained in the nasal passages and sinuses in the chinchilla, when the dye is administered in the prone position than when administered in the supine position but the converse is true for mice. Further, more dye is retained in the nasal passages and sinuses when administered to an alert chinchilla but again the converse is true for the mouse where an anesthetized state is preferred. In most cases, a divided dose permits more material to be retained in the nasal area.

[0016] The following table shows results for tests conducted with mice. Except as noted above, similar results occurred with tests conducted using chinchillas.

[0017] In the following tables “-” means that no dye is present, “traces” means that minimal amounts are present when examined with the unaided eye but that do not clearly show on photographs, “yes” and “+” mean dye is clearly visible, and “++” means heavy dye presence.

[0018] Table 1 shows the results for a control mouse treated with 10 μl of phosphate buffer solution (PBS) and no dye.

[0019] Table 2 shows the results for dye administered in various concentrations in a single dose with heavy anesthesia.

[0020] Table 3 shows the results for dye administered in various concentrations in a single dose with moderate anesthesia.

[0021] Table 4 shows the results for dye administered in a supine position at various concentrations in a single dose with heavy anesthesia.

[0022] Table 5 shows the results for dye administered in various concentrations in a single dose to alert animals.

[0023] Table 6 shows the results for dye administered at 30 μl concentration in a single dose to alert animals.

[0024] Table 7 shows the results for dye administered dropwise at 30 μl and 50 μl concentrations under heavy and moderate anesthesia.

[0025] Table 8 shows the results for dye administered in 5 μl and 2 μl increments showing reduced dye in the stomach at lower incremental doses than larger incremental doses and less the same summed quantity supplied in a single dose. Table 3 shows the results for dye administered in various concentrations in a single dose with moderate anesthesia.

[0026] Table 9 shows optimal divided dose conditions for the mouse where essentially no dye reached the stomach and very little dye reached the esophagus.

TABLE 1

Control

Position

Position

Time

At

Post

To

Esophagus

Trachea

Mouse

Dye

Deliv-

Deliv-

Sacri-

Nose

Nasal

Oral

Upper/

Intes-

Upper/

#

Volume

ery

ery

Anesthesia

fice

Skin

Cavity

Cavity

Larynx

Lower

Stomach

tine

Lower

Lung

Notes

1

0 μl

N/A

N/A

N/A

N/A

−

−

−

−

−/−

−

−

−/−

−

Control

(10 μl

mouse;

of

10 μl

PBS)

of PBS

total.

[0027]

TABLE 2
Dye Test, 200 μl of anesthesia (heavy)
		Position	Position
Mouse	Dye	At	After	Anesthesia	Time To	Nose	Nasal	Oral
#	Volume	Delivery	Delivery	Dose/Level	Sacrifice	Skin	Cavity	Cavity
2	10 μl	Upright	Not held	200 μl	60 min	Yes	Yes	Yes
				(heavy)
3	10 μL	Upright	Not held	200 μl	60 min	Yes	Yes	Yes
				(heavy)
4	20 μl	Upright	Not held	200 μl	60 min	Yes	Yes	Yes
				(heavy)
5	20 μl	Upright	Not held	200 μl	60 min	Yes	Yes	Yes
				(heavy)
6	30 μl	Upright	Not held	200 μl	60 min	Yes	Yes	Yes
				(heavy)
7	30 μl	Upright	Not held	200 μl	60 min	Yes	Yes	Yes
				(heavy)
8	40 μl	Upright	Not held	200 μl	60 min	Yes	Yes	Yes
				(heavy)
9	50 μl	Upright	Not held	200 μl	60 min	Yes	Yes	Yes
				(heavy)
10	50 μl	Upright	Not held	200 μl	60 min	Yes	Yes	Yes
				(heavy)
		Esophagus			Trachea
Mouse		Upper/			Upper/
#	Larynx	Lower	Stomach	Intestine	Lower	Lung	Notes
2	Yes	Traces/	Traces	−	−/−	−	Dye did not travel
		Traces					far
3	Yes	Yes/Yes	+	−	Traces/	−	Traces of dye were
					Traces		seen in the bronchial
							bronchial tubes and
							also appeared in the
							stomach
4	Yes	Yes/Yes	+	−	−/−	−	Dye appeared in
							stomach
5	Yes	Yes/Yes	+	−	−/−	−	Dye appeared in
							stomach
6	Yes	Yes/Yes	+	−	Yes/Yes	+	Dye was present in the
						Left	stomach and left lung
						Lung
						only
7	Yes	Yes/Yes	−	−	Yes/Yes	+	Dye was throughout
							esophagus and stopped
							just before entering
							the stomach. Dye was
							mostly in the lungs
8	Yes	Yes/Yes	+	Yes	Yes/Yes	+	There was a slight
						Right	presence of dye in the
						lung	right lung and none in
							the left lung
9	Yes	Yes/Yes	+	−	Yes/Yes	+	Dye was prominent in
							all examined areas
							except intestine
10	Yes	Yes/Yes	+	−	Yes/Yes	++	See above

[0028]

TABLE 3
Dye Test, 120 μl of anesthesia (moderate)
		Position	Position
Mouse	Dye	At	Post	Anesthesia	Time To	Nose	Nasal	Oral
#	Volume	Delivery	Delivery	Dose/Level	Sacrifice	Skin	Cavity	Cavity
11	30 μl	Upright	Not held	120 μl	60 min	Yes	Yes	Yes
				(Mod)
12	30 μl	Upright	Not held	120 μl	60 min	Yes	Yes	Yes
				(Mod)
13	50 μl	Upright	Not held	120 μl	60 min	Yes	Yes	Yes
				(Mod)
14	50 μl	Upright	Not held	120 μl	60 min	Yes	Yes	Yes
				(Mod)
		Esophagus			Trachea
Mouse		Upper/			Upper/
#	Larynx	Lower	Stomach	Intestine	Lower	Lung	Notes
11	Yes	Yes/Yes	++	−	Yes/Yes	Traces	Traces of dye in the
							lungs, and heavy in
							the stomach
12	Yes	Yes/Yes	+	−	Yes/Yes	Traces	Less presence of the
							dye in the stomach
							than #11
13	Yes	Yes/Yes	++	−	Yes/Yes	Traces	Traces of dye in the
							lungs and heavy in
							the stomach
14	Yes	Yes/Yes	++	Yes	Yes/	−	Traces seen in
					Traces		esophagus and
							trachea. Heavy in
							the stomach

[0029]

TABLE 4

Dye Test; Supine/Supine

Position

Position

Time

At

Post

Anesthesia

To

Nasal

Oral

Esophagus

Trachea

Mouse

Dye

Deliv-

Deliv-

Dose/

Sacri-

Nose

Cav-

Cav-

Upper/

Intes-

Upper/

#

Volume

ery

ery

Level

fice

Skin

ity

ity

Larynx

Lower

Stomach

tine

Lower

Lung

Notes

15

10 μl

Supine

Supine

200 μl

60

Yes

Yes

−

Yes

−/−

−

−

−/−

−

(heavy)

min

16

20 μl

Supine

Supine

200 μl

60

Yes

Yes

Yes

Yes

Traces/−

−

−

Traces/

−

Dark spot in

(heavy)

min

Traces

lungs was

blood

17

30 μl

Supine

Supine

200 μl

60

Yes

Yes

Yes

Yes

Yes/Yes

+

−

Yes/Yes

−

Oral delivery

(heavy)

min

18

30 μl

Supine

Supine

200 μl

60

Yes

Yes

Yes

Yes

Yes/Yes

−

−

Yes/Yes

−

Dye was

(heavy)

min

present in

the

esophagus

[0030]

TABLE 5
Varying Dye Volume; Alert Animals
		Position	Position
Mouse	Dye	At	Post	Anesthesia	Time To	Nose	Nasal	Oral
#	Volume	Delivery	Delivery	Dose/Level	Sacrifice	Skin	Cavity	Cavity
19	10 μl	Upright	Not held	0 μl (Alert)	60 min	−	Yes	Yes
20	10 μl	Upright	Not held	0 μl (Alert)	60 min	−	Yes	Yes
21	30 μl	Upright	Not held	0 μl (Alert)	60 min	Yes	Yes	Yes
22	30 μl	Upright	Not held	0 μl (Alert)	60 min	Yes	Yes	Yes
23	50 μl	Upright	Not held	0 μl (Alert)	60 min	Yes	Yes	Yes
24	50 μl	Upright	Not held	0 μl (Alert)	60 min	Yes	Yes	Yes
		Esophagus			Trachea
Mouse		Upper/			Upper/
#	Larynx	Lower	Stomach	Intestine	Lower	Lung	Notes
19	Yes	−/−	+	Yes	−.−	−	Relative to other
							mice this mouse did
							not struggle much
							during delivery into
							first nare but
							slightly struggled
							during delivery into
							second nare.
20	Yes	−/−	Traces	Yes	−/−	−	See above.
21	Yes	−/−	++	Yes	−/−	−	There was a great
							deal of gurgling and
							coughing of dye. A
							lot of dye appeared
							immediately in the
							mouth upon
							delivery.
22	Yes	−/−	+	Yes	−/−	−	See above.
23	Yes	Traces/	++	Yes	−/−	−	Mouse sneezed and
		Traces					spit up dye into
							mouth from nasal
							cavity. It was
							very difficult to
							administer all 50 μl.
24	Yes	−/−	+	Yes	−/−	Traces	See above. Dye was
							present in traces in
							bronchii.

[0031]

TABLE 6

Varying Position During Administration; Alert Animals

Position

Position

Time

At

Post

Anesthesia

To

Nasal

Oral

Esophagus

Trachea

Mouse

Dye

Deliv-

Deliv-

Dose/

Sacri-

Nose

Cav-

Cav-

Upper/

Stom-

Intes-

Upper/

#

Volume

ery

ery

Level

fice

Skin

ity

ity

Larynx

Lower

ach

tine

Lower

Lung

Notes

25

30 μl

Prone

Not

0 μl

60

−

Yes

Traces

Traces

−/−

+

Yes

−/−

Yes

Dye almost

held

(Alert)

min

completely

moved into

the intestine.

26

30 μl

Prone

Not

0 μl

60

−

Yes

Traces

Traces

−/−

++

Yes

−/−

−

Dye moved

held

(Alert)

min

through the

esophagus

completely

and was in

the stomach.

[0032]

TABLE 7
Drop-Wise Administration of Dye
		Position	Position
Mouse	Dye	At	Post	Anesthesia	Time To	Nose	Nasal	Oral
#	Volume	Delivery	Delivery	Dose/Level	Sacrifice	Skin	Cavity	Cavity
27	30 μl	Upright	Not held	200 μl	60 min	Yes	Yes	Yes
	1 drop/			(heavy)
	5 sec
	interval
28	50 μl;	Upright	Not held	120 μl	60 min	Yes	Yes	Yes
	5 μl/			(Mod)
	nare at
	30 sec
	interval
		Esophagus			Trachea
Mouse		Upper/			Upper/
#	Larynx	Lower	Stomach	Intestine	Lower	Lung	Notes
27	Yes	Yes/Yes	+	−	−/−	−	Drops were released
							and inhaled slowly
							pausing for five
							seconds in between.
							All dye moved down
							esophagus and into
							stomach.
28	Yes	Yes/Yes	+	−	−/−	−	Drops were released
							and inhaled, slowly
							paused for 30 seconds
							in between. All dye
							moved down
							esophagus into
							stomach. Animal was
							not fully asleep.

[0033]

TABLE 8
20-40 μl of Dye, Divided Doses (different intervals), 200 μl of anesthesia (heavy*),
Supine/Supine
		Position	Position
Mouse	Dye	At	Post	Anesthesia	Time To	Nose	Nasal	Oral
#	Volume	Delivery	Delivery	Dose/Level	Sacrifice	Skin	Cavity	Cavity
29	30 μl;	Supine	Supine	200 μl	60 min	Yes	Yes	−
	5 μl/			(heavy)
	nare at
	10 min
	interval
30	30 μl;	Supine	Supine	200 μl	60 min	Yes	Yes	Yes
	5 μl/			(heavy)
	nare at
	10 min
	interval
31	20 μl;	Supine	Supine	200 μl	60 min	Yes	Yes	−
	2 μl/			(heavy)
	nare at
	t = 0, 2,
	7, 9 and
	11 min
32	20 μl;	Supine	Supine	200 μl	60 min	Yes	Yes	Yes
	2 μl/			(heavy)
	nare at
	2 min
	interval
33	20 μl;	Supine	Supine	200 μl	60 min	Yes	Yes	Yes
	2 μl/			(heavy)
	nare at
	5 min
	interval
34	40 μl;	Supine	Supine	200 μl	60 min	Yes	Yes	Yes
	2 μl/			(heavy)
	nare at
	5 min
	interval
35	30 μl;	Supine	Supine	200 μl	60 min	Yes	Yes	Yes
	2 μl/			(Mod*)
	nare at
	5 min
	interval
36	30 μl;	Supine	Supine	200 μl	60 min	Yes	Yes	Yes
	2 μl/			(Mod*)
	nare at
	5 min
	interval
		Esophagus			Trachea
Mouse		Upper/			Upper/
#	Larynx	Lower	Stomach	Intestine	Lower	Lung	Notes
29	Yes	Yes/Yes	+	−	Yes/Yes	+
30	Yes	Yes/Yes	+	−	Yes/Yes	Traces
31	Yes	−/−	−	−	−/−	−	The 5 min interval
							between t = 2 and
							t = 7 min favored
							in maintenance
32	Yes	Yes/Yes	+	−	−/−	−
33	Yes	Yes/−	−	−	−/−	−
34	Yes	Yes/Yes	++	−	−/−	−	Mouse started to
							wake after about 32
							μl were
							administered.
35	Yes	Yes/Yes	+	−	−/−	−	Mouse was NOT
							heavily anesthetized
							at like others given
							the same amount of
							anesthesia. It began
							to wake and move
							after being given
							16 μl of dye.
36	Yes	Yes/Yes	+	−	−/−	−	See above
*Some mice, even when given 200 μl of Ketamine/Xylazine could not be considered heavily anesthetized because they behaved like moderately anesthetized animals.

[0034]

TABLE 9
Optimal Conditions for Intranasal Delivery and Maintenance
		Position	Position
Mouse	Dye	At	Post	Anesthesia	Time To	Nose	Nasal	Oral
#	Volume	Delivery	Delivery	Dose/Level	Sacrifice	Skin	Cavity	Cavity
37	30 μl;	Supine	Supine	200 μl	60 min	Yes	Yes	Yes
	2 μl/			(heavy)
	nare at
	5 min
	interval
38	30 μl;	Supine	Supine	400 μl	60 min	Yes	Yes	Yes
	2 μl/			(heavy)
	nare at
	5 min
	interval
39	30 μl;	Supine	Supine	350 μl	60 min	Yes	Yes	Yes
	2 μl/			(heavy)
	nare at
	5 min
	interval
		Esophagus			Trachea
Mouse		Upper/			Upper/
#	Larynx	Lower	Stomach	Intestine	Lower	Lung	Notes
37	Yes	Traces/−	−	−	−/−	−	A small isolated
							patch was found
							halfway down
							esophagus. No dye
							was in stomach,
							trachea or lungs.
							The dark spots on
							the lungs are blood
							clots.
38	Yes	−/−	−	−	−/−	−	Animal needed
							additional anesthetic
							during dye adminis-
							tration in order to
							stay heavily
							anesthetized until
							dye was completely
							administered.
39	Yes	Traces/−	−	−	Traces/−	−	Animal needed
							additional anesthetic
							during dye adminis-
							tration in order to
							stay heavily
							anesthetized until
							dye was completely
							administered.

Claims

1. A method for nasal application of a medicinal substance which comprises applying the substance through the nose in a maximum amount that is insufficient to immediately stimulate an excretory response that would clear a significant portion of the substance from nasal and sinus passages and within a time period of less than one hour, and repeating the application of the substance, through the nose in a maximum amount that is insufficient to immediately stimulate an excretory response that would clear a significant portion of the substance from nasal and sinus passages, at least once and at a minimum a sufficient number of times to provide an effective total dose of the substance.

2. The method of claim 1 where the application is repeated a sufficient number of times within the hour to maximize the total dose without stimulating an excretory response that would clear a significant portion of the substance from nasal and sinus passages.

3. The method of claim 1 where the substance is a nasally absorbable medicine.

4. The method of claim 1 where the substance is a nasally active medicine.

5. The method of claim 1 where the substance is selected from the group consisting of vaccines, antigens, epitopes, adjuvants, viral vectors, bacterial vectors, immune modulators, delivery vehicles, and drugs including antibiotics, hormones, antibodies, anti-inflammatories, antipyretics, antispasmotics, anesthetics, chemotherapeutic agents, sedatives, analgesics, vasodialators, and vasoconstrictors.

6. The method of claim 1 where the number of applications is from 3 to about 15 applications within the hour.

7. The method of claim 1 where the mammal is a supine mouse.

8. The method of claim 1 where the mammal is a prone chinchilla.

9. The method of claim 1 where the mammal is a human.

10. The method of claim 1 where the medical substance is a vaccine.

11. The method of claim 10 where the vaccine is a vaccine for non-typeable haemophilus influenzae.

12. The method of claim 11 where the vaccine contains an epitope of P6 protein of haemophilus influenzae.

13. The method of claim 11 where the vaccine contains an epitope of P5 protein of haemophilus influenzae.

14. The method of claim 11 where the vaccine is a vaccine against hepatitis B.