Compounds and adjuvant preparations useful for pneumococcal vaccines

Abstract

The present invention relates to novel compounds and formulations. Additionally, the present invention relates to novel compounds and formulations useful for pneumococcal and pneumococcal conjugate vaccines. More specifically, the present invention relates to compositions comprising a pneumococcal conjugate and one or more compounds of Formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or pharmaceutically acceptable salts thereof, formulated as a stable nanoemulsion (referred to herein as an "SNE adjuvant composition" or "SNE").

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the benefits of U.S. Provisional Patent Application No. 63 / 503,021, filed on 18 May 2023, the entirety of which is incorporated herein by reference. [Background technology]

[0002] Pneumococcal disease is caused by infection with the bacterium Streptococcus pneumoniae (Pneumococcus). Different serotypes of Pneumococcus are known to cause different manifestations of the disease, and infection can cause a range of symptoms from ear and sinus infections to pneumonia and bloodstream infections. Pneumococcal disease has a high associated morbidity and mortality rate worldwide, particularly among the elderly and children. Currently, 100 capsular polysaccharides (some of which are used to define or specify particular serotypes) have been identified (Ganaie, F. et al. (2020) Clinical Science and Epidemiology, Vol. 11, Issue 3, pages 1-15). These serotypes are distinguished by their chemical structure, serological response, and other associated genetic mutations.

[0003] In 1983, PNEUMOVAX® (Merck & Co., Inc., Loway, New Jersey, USA), a 23-valent pneumococcal vaccine (PV), was approved in the United States. This vaccine demonstrated reduced immunogenicity in infants due to a T-cell independent response. To address this problem, pneumococcal conjugate vaccines (PCVs) were developed, particularly in infants. By covalently attaching a polysaccharide to a carrier protein, the immunogenic response became T-cell dependent. In 2000, PREVNAR® (Pfizer Inc., Philadelphia, Pennsylvania, USA), a 7-valent PCV, was approved in the United States. In 2010, PREVNAR13® (Pfizer Inc.), a 13-valent PCV, was approved in the United States. In 2021, VAXNEUVANCE® (Merck & Co., Inc., Loway, New Jersey, USA), a 15-valent PCV, and PREVNAR20® (Pfizer Inc.), a 20-valent PCV, were approved in the United States. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Ganaie,F.et al.(2020)Clinical Science and Epidemiology,Vol.11,Issue 3,pages 1-15 [Overview of the project] [Problems that the invention aims to solve]

[0005] Approved PCVs currently utilize aluminum-containing derivatives as adjuvants to enhance immunogenicity. While aluminum adjuvants increase the immunogenic response from baseline, it is unclear whether this response is sufficient for higher-tidal PCVs, particularly in infants. Therefore, there is a need to identify other adjuvants that can increase the immunogenicity of polyvalent PCVs beyond the current aluminum adjuvant standard. [Means for solving the problem]

[0006] The present invention relates to formulations and compositions comprising (i) one or more Streptococcus pneumoniae (S. pneumoniae) antigens, (ii) one or more compounds of formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or (one or more) pharmaceutically acceptable salts thereof, and (iii) a pharmaceutically acceptable carrier, wherein one or more Streptococcus pneumoniae antigens can induce an immune response to one or more serotypes of Streptococcus pneumoniae.

[0007] The present invention also relates to formulations and compositions comprising: (i) one or more Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of which contains a polysaccharide of a specific Streptococcus pneumoniae serotype conjugate to a carrier protein, and the carrier protein being CRM197; (ii) one or more compounds of formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or (one or more) pharmaceutically acceptable salts thereof; and (iii) a pharmaceutically acceptable carrier. Furthermore, the present invention relates to the use of a formulation or composition for the prevention of pneumococcal disease or for the prevention or reduction of the possibility of infection by Streptococcus pneumoniae, comprising: (i) one or more Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of which contains a polysaccharide of a specific Streptococcus pneumoniae serotype conjugate to a carrier protein, and the carrier protein being CRM197; (ii) one or more compounds of formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or (one or more) pharmaceutically acceptable salts thereof; and (iii) a pharmaceutically acceptable carrier.

[0008] The present invention also relates to formulations and compositions comprising (i) one or more Streptococcus pneumoniae antigens, (ii) one or more compounds of formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) one or more sorbitan-based surfactants, and (iv) one or more terpenes. In some embodiments, one or more Streptococcus pneumoniae antigens are Streptococcus pneumoniae polysaccharide carrier protein conjugates, each of which contains a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein. In some embodiments, the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.

[0009] The present invention also relates to formulations and compositions comprising (i) one or more Streptococcus pneumoniae antigens, (ii) one or more compounds of formulas I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) sorbitan trioleate (SPAN-85), (iv) polysorbate-20 (PS-20) or polysorbate-80 (PS-80), and (v) squalene. In some embodiments, one or more Streptococcus pneumoniae antigens are Streptococcus pneumoniae polysaccharide carrier protein conjugates, each conjugate containing a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein. In some embodiments, the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197. In some embodiments, the Streptococcus pneumoniae antigen is a polysaccharide carrier protein conjugate, where the polysaccharide consists of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.

[0010] The present invention also relates to a composition comprising a pneumococcal conjugate disclosed herein and a stable nanoemulsion "SNE". As an example, a particular SNE comprises N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)pipradin-1-yl)-5-oxopentyl)stearamide, also known as "compound A-1". This particular SNE is called compound A-1-SNE and comprises i) compound A-1, ii) sorbitan trioleate (SPAN-85), (iii) polysorbate-20 (PS-20) or polysorbate-80 (PS-80), and (iv) squalene. As another example, certain SNEs include N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, also known as "compound B-1". This particular SNE is called compound B-1-SNE and includes i) compound B-1, ii) SPAN-85, (iii) PS-20 or PS-80, and (iv) squalene. As yet another example, certain SNEs include N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, also known as "compound C-4". This particular SNE is called compound C-4-SNE and comprises i) compound C-4, ii) SPAN-85, (iii) PS-20 or PS-80, and (iv) squalene.

[0011] The present invention also relates to an immunogenic composition comprising pneumococcal polysaccharides or pneumococcal conjugates and SNEs as disclosed herein.

[0012] The present invention also relates to a method for treating or preventing pneumococcal disease in patients in need by administering the immunogenic composition of the present invention to the patient.

[0013] The present invention also relates to a method for preventing or reducing the likelihood of infection by one or more serotypes of Streptococcus pneumoniae, comprising administering the immunogenic composition of the present invention to a patient in need thereof. [Brief explanation of the drawing]

[0014] [Figure 1] (Example 10) This figure shows the OD640 readouts after incubation of human TLR7 (upper panel) or TLR8 (lower panel) expressing HEK-Blue® 293 cells (InvivoGen) with different concentrations of compound B-1-SNE. The Y axis represents the total concentration of compound B-1-SNE. [Figure 2] (Example 11) This figure shows the anti-6B IgG titers before immunization (day 0, pooled serum), post-dose 1 (day 21), post-dose 2 (day 42), and post-dose 3 (day 70) after immunizing mice with the formulations listed in Table 7. Error bars are geometric mean with 95% confidence intervals. Transformed data analyzed by one-way ANOVA with Dunnett post-hoc test, *p<0.05. [Figure 3] (Example 11) This figure shows the anti-6B OPA titers from pooled serum of mice immunized with the formulations listed in Table 7, before immunization (day 0), after dose 1 (day 21), after dose 2 (day 42), and after dose 3 (day 70). [Figure 4] (Example 12) This figure shows the ratio of serotype-specific IgG titers in infant rhesus monkeys after immunization with PCV24 containing compound B-1-SNE (compound B-1 at a 150 μg dose), PCV24 containing compound B-1-SNE (compound B-1 at a 50 μg dose), and PCV24 containing compound B-1-SNE (compound B-1 at a 15 μg dose), compared to immunization with PCV24 formulated without adjuvant 3 doses post-dose. Data for serotypes 6C and 15B are included to assess cross-reactivity. [Figure 5](Example 12) A diagram showing the ratio of serotype-specific IgG titers in infant cynomolgus monkeys after immunization with PCV24 having compound B-1-SNE (150 μg dose of compound B-1), PCV24 having compound B-1-SNE (50 μg of compound B-1), and PCV24 having compound B-1-SNE (15 μg of compound B-1), compared with the immunization of PCV24 formulated with APA 3 days after dosing. Data for serotypes 6C and 15B are included to evaluate cross-reactivity. [Figure 6] (Example 12) A diagram showing the ratio of serotype-specific IgG titers in infant cynomolgus monkeys after immunization with PCV24 having compound B-1-SNE (150 μg dose of compound B-1), PCV24 having compound B-1-SNE (50 μg of compound B-1), and PCV24 having compound B-1-SNE (15 μg of compound B-1), compared with the immunization of PCV20 3 days after dosing. [Figure 7] (Example 13) A diagram showing the ratio of serotype-specific IgG titers in infant cynomolgus monkeys after immunization with PCV24 having compound B-1-SNE (100 μg dose of compound B-1), PCV24 having compound B-1-SNE (10 μg of compound B-1), PCV24 having compound B-1-SNE (1 μg of compound B-1), and PCV24 having compound B-1-SNE (0.1 μg of compound B-1), compared with the immunization of PCV20 3 days after dosing. [Figure 8] (Example 14) A diagram showing the ratio of serotype-specific IgG titers in mice after immunization with PCV26 having compound B-1-SNE (10 μg dose of compound B-1), compared with the immunization of PCV26 formulated with APA 2 days after dosing. Data for serotypes 6C and 15B are included to evaluate cross-reactivity. [Figure 9] (Example 14) A diagram showing the IgG2a / IgG1 titer ratio of serotype-specific antibodies in mice after immunization with PCV26 having compound B-1-SNE (10 μg dose of compound B-1) or PCV26 formulated with APA 3 days after dosing. Responses from three representative serotypes 18C, 19F, and 24F are shown (from left to right). [Figure 10A](Example 14) This is a diagram showing that mice immunized with PCV26 formulated with compound B-1-SNE (compound B-1 at a dose of 10 μg) or PCV26 formulated with APA are protected from intratracheal challenge with Streptococcus pneumoniae 24F. Bacteremia was quantified 24 hours after infection with serotype 24F bacteria and shown as log CFU / mL. [Figure 10B] (Example 14) This is a diagram showing that mice immunized with PCV26 formulated with compound B-1-SNE (compound B-1 at a dose of 10 μg) or PCV26 formulated with APA are protected from intratracheal challenge with Streptococcus pneumoniae 24F. Survival rate was evaluated over approximately 240 hours after infection with serotype 24F bacteria.

Modes for Carrying Out the Invention

[0015] The present invention relates to one or more Streptococcus pneumoniae antigens and a compound having the structure described in Formula I:

Chemical Formula

Chemical formula

Chemical formula

[0016] The present invention involves one or more Streptococcus pneumoniae antigens and a compound having the structure described in formula Ia: [ka] (In the formula, R' and R'' are independently selected from H, (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl, and the (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine, or R' and R'' together with the nitrogen to which they are bonded to form a (C3-C6) heterocycloalkyl, and the (C3-C6) heterocycloalkyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. R bEach occurrence of -O(C1-C4)alkyl, and said -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. A is

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0017] The present invention involves one or more Streptococcus pneumoniae antigens and a compound having the structure described in Formula II: [ka] (In the formula, R 1 is a (C1-C6)alkyl group, and the (C1-C6)alkyl group may be substituted with 1 to 4 substituents independently selected from -OH and -O(CH3). R 2 is H, methyl, or -O(CH3), R 3 Each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH and -O(CH3). R 4 Each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH and -O(CH3). R 5 teeth, [ka] And, R 6 (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) Selected from alkynyl, and the above (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 The alkynyl group may be substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. Each occurrence of n is 4. The present invention provides a composition comprising either or a pharmaceutically acceptable salt thereof.

[0018] The present invention involves one or more Streptococcus pneumoniae antigens and a compound having the structure described in formula IIa: [ka] (In the formula, R 1 is butyl, and the butyl may be substituted with one or two -OH groups. R 3 Each occurrence is independently either H or -O(CH3), R 5 teeth, [ka] And, R 6 is, (C 10 -C 20 ) alkyl, (C 10 -C 20 ) Alkenyl, and (C 10 -C 20 (Selected from Alkinil) The present invention provides a composition comprising either or a pharmaceutically acceptable salt thereof.

[0019] The present invention involves one or more Streptococcus pneumoniae antigens and a compound having the structure described in Formula III: [ka] (In the formula, R 1 is a (C1-C6)alkyl group, and the (C1-C6)alkyl group may be substituted with 1 to 4 substituents independently selected from -OH and -O(CH3). R 2 is H, methyl, or -O(CH3), R 3 Each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH and -O(CH3). R 4 (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) Selected from alkynyl, and the above (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 The alkynyl group may be substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. n is 4. The present invention provides a composition comprising either or a pharmaceutically acceptable salt thereof.

[0020] The present invention involves one or more Streptococcus pneumoniae antigens and a compound having the structure described in formula IIIa: [ka] During the ceremony, (R 1 is butyl, and the butyl may be substituted with one or two -OH groups. R 3 Each occurrence is independently either H or -O(CH3), R 4 is, (C 10 -C 20 ) alkyl, (C 10 -C 20 ) Alkenyl, and (C 10 -C 20 (Selected from Alkinil) The present invention provides a composition comprising either or a pharmaceutically acceptable salt thereof.

[0021] The present invention involves one or more Streptococcus pneumoniae antigens and a compound having the structure described in Formula IV: [ka] During the ceremony, (R 1 is a (C1-C6)alkyl group, and the (C1-C6)alkyl group may be substituted with 1 to 4 substituents independently selected from -OH and -O(CH3). R 2 is H, methyl, or -O(CH3), R 3 Each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, and -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH and -O(CH3). R 4 Each occurrence is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) independently selected from alkynyl, and the (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20The alkynyl group may be substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, or fluorine. n is 4. The present invention provides a composition comprising either or a pharmaceutically acceptable salt thereof.

[0022] The present invention involves one or more Streptococcus pneumoniae antigens and a compound having the structure described in formula IVa: [ka] (In the formula, R 1 is butyl, and the butyl may be substituted with one or two -OH groups. R 3 Each occurrence is independently either H or -O(CH3), R 4 Each occurrence is (C 10 -C 20 ) alkyl, (C 10 -C 20 ) Alkenyl, and (C 10 -C 20 (Selected independently of Alkinnil) The present invention provides a composition comprising either or a pharmaceutically acceptable salt thereof.

[0023] This invention relates to one or more Streptococcus pneumoniae antigens and one or more of the following compounds: Also known as compound A-1, (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl) stearamide, Also known as compound A-2, (S)-N-(5-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, Also known as compound A-3, (S)-1-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-yl)octadecane-1-one, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, also known as compound B-1, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-yl)-5-oxopentyl)tetradecaneamide, also known as compound B-2, Also known as compound B-3, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)oleamide, Also known as compound B-4, (9Z,12Z)-N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)octadeca-9,12-dienamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepan-1-yl)-5-oxopentyl)stearamide, also known as compound B-5, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperidine-1-yl)-5-oxopentyl)stearamide, also known as compound B-6, N-(5-(3-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)azetidine-1-yl)-5-oxopentyl)stearamide, also known as compound B-7, Compound B-8, also known as 1-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperidine-4-carboxamide, Also known as compound B-9, (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-octadecylcyclobutane-1-carboxamide (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-hexadecylcyclobutane-1-carboxamide, also known as compound B-10. N-(3-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-3-oxopropyl)stearamide, also known as compound B-11, N-(7-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-7-oxoheptyl)stearamide, also known as compound B-12, Also known as compound B-13, N-(3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)cyclobutyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-4,4-dimethyl-5-oxopentyl)stearamide, also known as compound B-14, N-(6-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-yl)-2-methyl-6-oxohexane-2-yl)stearamide, also known as compound B-15, Also known as compound B-16, 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecyloxy)pentan-1-one, Compound B-17, also known as 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecylamino)pentan-1-one, N-(5-(4-(4-((5-amino-7-(butylamino)-3-methyl-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, also known as compound B-18, Also known as compound C-1, (9Z,12Z)-N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)octadeca-9,12-dienamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)tetradecaneamide, also known as compound C-2, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)oleamide, also known as compound C-3, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, also known as compound C-4, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)-4-oxobutyl)stearamide, also known as compound C-5, Also known as compound D-1, (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)carbamate, Also known as compound D-2, 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperazine-1-carboxamide, 3-stearamidopropyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-carboxylate, also known as compound D-3, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-6,6,6-trifluorohexyl)stearamide, also known as compound D-4, N-(4-((4-((7-(butylamino)-5-hydroxy-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, also known as compound D-5, and Also known as compound D-6, (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-yl)-5-oxopentyl)carbamate, The present invention provides a composition comprising either or a pharmaceutically acceptable salt thereof.

[0024] The present invention also provides a composition comprising one or more Streptococcus pneumoniae antigens and a stable nanoemulsion, wherein the stable nanoemulsion comprises (i) one or more compounds having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or (one or more) pharmaceutically acceptable salts thereof, ii) one or more emulsifiers, and iii) one or more terpenes.

[0025] The present invention also provides a composition comprising one or more Streptococcus pneumoniae antigens and a stable nanoemulsion, wherein the stable nanoemulsion comprises (i) one or more compounds having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or (one or more) pharmaceutically acceptable salts thereof, ii) one or more sorbitan-based surfactants, and iii) one or more terpenes.

[0026] The present invention also provides a composition comprising one or more Streptococcus pneumoniae antigens and a stable nanoemulsion, wherein the stable nanoemulsion comprises (i) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (ii) SPAN-85, (iii) PS-20 or PS-80, and (iv) squalene.

[0027] The present invention also provides a composition comprising one or more Streptococcus pneumoniae antigens and a stable nanoemulsion, wherein the stable nanoemulsion comprises (i) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof, (ii) SPAN-85, (iii) PS-20 or PS-80, and (iv) squalene.

[0028] The present invention also provides a pharmaceutical composition comprising (i) at least one Streptococcus pneumoniae (S. pneumoniae) polysaccharide, (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, (v) squalene, and (vi) a pharmaceutically acceptable carrier.

[0029] The present invention also provides a pharmaceutical composition comprising (i) at least one Streptococcus pneumoniae (S. pneumoniae) polysaccharide, (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl) stearamide, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, (v) squalene, and (vi) a pharmaceutically acceptable carrier.

[0030] The present invention also provides an immunogenic composition comprising (i) at least one Streptococcus pneumoniae polysaccharide, (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, (v) squalene, and (vi) a pharmaceutically acceptable carrier.

[0031] The present invention also provides an immunogenic composition comprising (i) at least one Streptococcus pneumoniae polysaccharide, (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, (v) squalene, and (vi) a pharmaceutically acceptable carrier.

[0032] The present invention also provides a single-dose vaccine composition comprising (i) at least one Streptococcus pneumoniae polysaccharide, (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, and (iv) PS-20 or PS-80, wherein a single dose of the vaccine composition is sufficient to induce a desired immune response to at least one Streptococcus pneumoniae polysaccharide.

[0033] The present invention also provides a single-dose vaccine composition comprising (i) at least one Streptococcus pneumoniae polysaccharide, (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, (v) squalene, and (vi) a pharmaceutically acceptable carrier, wherein a single dose of the vaccine composition is sufficient to induce a desired immune response to at least one Streptococcus pneumoniae polysaccharide.

[0034] The present invention also provides a method for treating or preventing pneumococcal disease in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0035] The present invention also provides a method for treating or preventing pneumococcal disease in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide; (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0036] The present invention also provides a pharmaceutical composition comprising (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, and (v) squalene.

[0037] The present invention also provides a pharmaceutical composition comprising (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, (v) squalene, and (vi) a pharmaceutically acceptable carrier.

[0038] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a specific Streptococcus pneumoniae polysaccharide conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 1 9A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22 The present invention provides a pharmaceutical composition comprising: (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a group of serotypes selected from F, 23A, 23B, 23F, 24F, 33F, and 35B, with the carrier protein being CRM197; (ii) a compound having the structure described in formulas I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0039] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a specific Streptococcus pneumoniae polysaccharide conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A c) 1, 3, 4, 5, 6A, 6B, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 2 2F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23 The present invention provides a pharmaceutical composition comprising: (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a group of serotypes selected from F, 24F, 33F, and 35B, with the carrier protein being CRM197; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0040] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15 A) De-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 3 From the group of serotypes selected from 3F and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B The present invention provides a pharmaceutical composition comprising: (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0041] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, o -Acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35 B, e) consists of a group of serotypes selected from 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) consists of a group of serotypes selected from 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is C The present invention provides a pharmaceutical composition comprising: (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0042] The present invention also provides an immunogenic composition comprising (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, and (v) squalene.

[0043] The present invention also provides an immunogenic composition comprising (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, (v) squalene, and (vi) a pharmaceutically acceptable carrier.

[0044] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a specific Streptococcus pneumoniae polysaccharide conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19 A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, The present invention provides an immunogenic composition comprising: (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a group of serotypes selected from 23A, 23B, 23F, 24F, 33F, and 35B, with the carrier protein being CRM197; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0045] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, as well as f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 2 The present invention provides an immunogenic composition comprising: (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a group of serotypes selected from 4F, 33F, and 35B, with the carrier protein being CRM197; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0046] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15 A) De-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33 F and 35B, e) from a group of serotypes selected from 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) from a group of serotypes selected from 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. The present invention provides an immunogenic composition comprising: (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0047] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, o -Acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B e) consists of a group of serotypes selected from 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) consists of a group of serotypes selected from 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM The present invention provides an immunogenic composition comprising: (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0048] The present invention also provides a single-dose vaccine composition comprising (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, and (v) squalene, wherein a single dose of the vaccine composition is sufficient to induce a desired immune response against at least one polysaccharide carrier protein conjugate.

[0049] The present invention also provides a single-dose vaccine composition comprising (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, (v) squalene, and (vi) a pharmaceutically acceptable carrier.

[0050] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A , de-O-acetylation 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) consists of a group of serotypes selected from 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and The present invention provides a single-dose vaccine composition comprising: (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate whose somatic protein is CRM197; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene, wherein a single dose of the vaccine composition is sufficient to induce a desired immune response against the polysaccharide carrier protein conjugate.

[0051] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, deoxygenated O- Acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e ) Consists of a group of serotypes selected from 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, wherein the carrier protein is CRM197 The present invention provides a single-dose vaccine composition comprising: (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier, wherein a single dose of the vaccine composition is sufficient to induce a desired immune response against the polysaccharide carrier protein conjugate.

[0052] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a specific Streptococcus pneumoniae serotype polysaccharide conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18 c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, as well as e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and e) 1, 3, 4, 5, 6A, 6B, 7F Streptococcus pneumoniae polysaccharide carrier protein conjugates consisting of a group of serotypes selected from 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, with the carrier protein being CRM197. The present invention provides a single-dose vaccine composition comprising (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof, (iii) SPAN-85, (iv) PS-20 or PS-80, and (v) squalene, wherein a single dose of the vaccine composition is sufficient to induce a desired immune response against a polysaccharide carrier protein conjugate.

[0053] The present invention also relates to (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a specific Streptococcus pneumoniae serotype polysaccharide conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 1 c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 1 (ii)N-(5-(4-( The present invention provides a single-dose vaccine composition comprising (iii) SPAN-85, (iv) PS-20 or PS-80, (v) squalene, and (vi) a pharmaceutically acceptable carrier, wherein a single dose of the vaccine composition is sufficient to induce a desired immune response against a polysaccharide carrier protein conjugate.

[0054] The present invention also provides a method for treating or preventing pneumococcal disease in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0055] The present invention also provides a method for treating or preventing pneumococcal disease in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0056] The present invention also relates to a method for treating or preventing pneumococcal disease in human patients, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16 F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, as well as f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A The present invention provides a method comprising administering to a patient a pharmaceutical composition comprising: (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a group of serotypes selected from 23B, 23F, 24F, 33F, and 35B, with the carrier protein being CRM197; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0057] The present invention also provides a method for treating or preventing pneumococcal disease in human patients, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19 A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F The present invention provides a method comprising administering to a patient a pharmaceutical composition comprising: (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a group of serotypes selected from 33F and 35B, the carrier protein being CRM197; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0058] The present invention also relates to a method for treating or preventing pneumococcal disease in human patients, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 2 It consists of a group of serotypes selected from 3F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and The present invention provides a method comprising administering to a patient a pharmaceutical composition comprising: (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0059] The present invention also provides a method for treating or preventing pneumococcal disease in human patients, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10 A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33 The group consists of serotypes selected from F and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197 The present invention provides a method comprising administering to a patient a pharmaceutical composition comprising: (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0060] The present invention also provides a method for preventing or reducing the likelihood of pneumococcal infection in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0061] The present invention also provides a method for preventing or reducing the likelihood of pneumococcal infection in a human patient, comprising administering to the patient a pharmaceutical composition comprising: (i) at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0062] The present invention also provides a method for preventing or reducing the likelihood of pneumococcal infection in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which contains a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15 B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, as well as f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 2 The present invention provides a method comprising administering to a patient a pharmaceutical composition comprising: (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a group of serotypes selected from 3A, 23B, 23F, 24F, 33F, and 35B, with the carrier protein being CRM197; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0063] The present invention also provides a method for preventing or reducing the likelihood of pneumococcal infection in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4 c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, as well as f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24 The present invention provides a method comprising administering to a patient a pharmaceutical composition comprising: (ii) a Streptococcus pneumoniae polysaccharide carrier protein conjugate consisting of a group of serotypes selected from F, 33F, and 35B, with the carrier protein being CRM197; (ii) a compound having the structure described in formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0064] The present invention also provides a method for preventing or reducing the likelihood of pneumococcal infection in a human patient, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23 The group consists of serotypes selected from B, 23F, 24F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. The present invention provides a method comprising administering to a patient a pharmaceutical composition comprising: (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; and (v) squalene.

[0065] The present invention also provides a method for preventing or reducing the likelihood of pneumococcal infection in human patients, comprising: (i) a Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which comprises a specific Streptococcus pneumoniae serotype polysaccharide conjugated to a carrier protein, wherein the Streptococcus pneumoniae polysaccharide is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B; b) 1, 3, 4, 5, 6A, 6B, 7F, 8 c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B, d) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24 The group consists of serotypes selected from F, 33F, and 35B, e) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and f) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM1 The present invention provides a method comprising administering to a patient a pharmaceutical composition comprising: (ii) N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide or a pharmaceutically acceptable salt thereof; (iii) SPAN-85; (iv) PS-20 or PS-80; (v) squalene; and (vi) a pharmaceutically acceptable carrier.

[0066] In specific embodiments of the above-mentioned i) pharmaceutical composition, ii) immunogenic composition, and iii) single-dose vaccine composition, the composition comprises more than 20(20) different Streptococcus pneumoniae polysaccharide carrier protein conjugates. For example, the present invention provides a pharmaceutical composition comprising a Streptococcus pneumoniae polysaccharide carrier protein conjugate comprising a Streptococcus pneumoniae polysaccharide carrier protein conjugate comprising a specific Streptococcus pneumoniae serotype polysaccharide conjugate, wherein the Streptococcus pneumoniae polysaccharide conjugate comprises a group of serotypes selected from more than 20(20) different Streptococcus pneumoniae polysaccharides.

[0067] In the specific embodiments described above, the serotype group is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, or b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A , de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, or c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

[0068] In the specific embodiments described above, the serotype group is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, or b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated These are 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, or c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.

[0069] In specific embodiments of the above-mentioned i) pharmaceutical composition, ii) immunogenic composition, and iii) single-dose vaccine composition, the composition comprises more than 25(25) different Streptococcus pneumoniae polysaccharide carrier protein conjugates. For example, the present invention provides a pharmaceutical composition comprising a Streptococcus pneumoniae polysaccharide carrier protein conjugate comprising a Streptococcus pneumoniae polysaccharide carrier protein conjugate comprising a specific Streptococcus pneumoniae serotype polysaccharide conjugate, wherein the Streptococcus pneumoniae polysaccharide conjugate comprises a group of serotypes selected from more than 25(25) different Streptococcus pneumoniae polysaccharides.

[0070] In the specific embodiments described above, the serotype group is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, or b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A , de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, or c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

[0071] In the specific embodiments described above, the serotype group is a) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, or b) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated These are 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, or c) 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197.

[0072] In some embodiments of the above-mentioned i) pharmaceutical composition, ii) immunogenic composition, and iii) single-dose vaccine composition, the concentration of the compound is 0.01 μg / mL to 1000 μg / mL or 0.1 μg / mL to 100 μg / mL or 80 μg / mL or 16 μg / mL or 4 μg / mL.

[0073] In some embodiments of the above-mentioned i) pharmaceutical composition, ii) immunogenic composition, and iii) single-dose vaccine composition, the concentration of SPAN-85 is 0.001 mg / mL to 100 mg / mL or 0.01 mg / mL to 50 mg / mL or 0.1 mg / mL to 10 mg / mL.

[0074] In some embodiments of the above-mentioned i) pharmaceutical composition, ii) immunogenic composition, and iii) single-dose vaccine composition, the concentration of PS-20 or PS-80 is 0.001 mg / mL to 100 mg / mL or 0.01 mg / mL to 50 mg / mL or 0.1 mg / mL to 10 mg / mL.

[0075] In some embodiments of the above-mentioned i) pharmaceutical composition, ii) immunogenic composition, and iii) single-dose vaccine composition, the concentration of squalene is 0.01 mg / mL to 100 mg / mL or 0.02 mg / mL to 20 mg / mL or 1 mg / mL to 20 mg / mL.

[0076] In some embodiments of the above-mentioned i) pharmaceutical composition, ii) immunogenic composition, and ii) single-dose vaccine composition, the concentration of the compound is 80 μg / mL, the concentration of SPAN-85 is 4.8 mg / mL, the concentration of PS-20 or PS-80 is 4.8 mg / mL, and the concentration of squalene is 16 mg / mL.

[0077] i) pharmaceutical composition, ii) immunogenic composition, and ii) single-dose vaccine composition described above, and iv) in some embodiments of the single-dose vaccine composition described above, the concentration of the compound is 16 μg / mL, the concentration of SPAN-85 is 4.8 mg / mL, the concentration of PS-20 or PS-80 is 4.8 mg / mL, and the concentration of squalene is 16 mg / mL.

[0078] i) pharmaceutical composition, ii) immunogenic composition, and ii) single-dose vaccine composition described above, and iv) in some embodiments of the single-dose vaccine composition described above, the concentration of the compound is 4 μg / mL, the concentration of SPAN-85 is 4.8 mg / mL, the concentration of PS-20 or PS-80 is 4.8 mg / mL, and the concentration of squalene is 16 mg / mL.

[0079] In some embodiments of the above-mentioned i) pharmaceutical composition, ii) immunogenic composition, and iii) single-dose vaccine composition, the composition further comprises L-met and EDTA.

[0080] Abbreviations and definitions The following abbreviations are used herein.

[0081] [Table 1] TIFF0007872897000018.tif230151TIFF0007872897000019.tif227136TIFF0007872897000020.tif204152

[0082] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural references unless the context clearly indicates otherwise.

[0083] As used herein, the term “about” means, when used herein in relation to a value, the same as the referenced value, or, in the context, similar to the referenced value. In general, a person skilled in the art familiar with the context will understand the absolute amount and / or relative degree of the difference encompassed by “about” in that context. For example, in some embodiments, the term “about” may encompass a range of values ​​of 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the referenced value.

[0084] As defined herein, “adjuvant” means a compound or compound formulation or composition that helps enhance the immunogenicity of the compositions of the present invention. Adjuvants may: i) enhance an immune response to an antigen that is weakly immunogenic when administered alone (e.g., Streptococcus pneumoniae polysaccharide), for example, by not inducing an antibody titer or cell-mediated immune response, or by inducing a weak antibody titer or cell-mediated immune response; ii) increase the antibody titer against the antigen, and / or iii) decrease the dose of the antigen that is effective in achieving an immune response in an individual.

[0085] As used herein, the term “administration” refers to the act of providing an active agent, composition, or formulation to the human body. Exemplary routes of administration to the human body may include the eyes (eye drops), mouth (oral), skin (transdermal), nose (transnasal), lungs (inhalation), rectum, vagina, oral mucosa (cheek), ear, and injection (e.g., intravenous (IV), subcutaneous, intratumoral, intraperitoneal, intramuscular (IM), intradermal (ID), etc.).

[0086] As used herein, “drug” means any particle, compound, molecule, or entity of any chemical class.

[0087] As used herein, the term “alkyl” refers to a linear, cyclic, or branched saturated aliphatic hydrocarbon having a specified number of carbon atoms. A numerical range may be given to indicate the total chain length. For example, C1-C6 alkyls have a chain length of 1 to 6 atoms. Alkyl groups may be the same or different and may be substituted with one or more “substituents” as defined herein below. Unless otherwise indicated, alkyl groups are unsubstituted.

[0088] As used herein, the term “alkenyl” means a linear, cyclic, or branched unsaturated aliphatic hydrocarbon having a specified number of carbon atoms, including but not limited to dienes, trienes, and tetraenes. The alkenyl group may be the same or different and may be substituted with one or more “substituents” as defined herein below. Unless otherwise indicated, the alkenyl group is unsubstituted.

[0089] As used herein, the term “alkynyl” means a linear, cyclic, or branched unsaturated aliphatic hydrocarbon having a specified number of carbon atoms, including but not limited to dienes, trienes, and tetraenes. The alkynyl group may be the same or different and may be substituted with one or more “substituents” as defined herein below. Unless otherwise indicated, the alkynyl group is unsubstituted.

[0090] As used herein, the term “antigen” refers to any antigen capable of generating one or more immune responses. An antigen may be a protein, peptide, or polypeptide. In certain embodiments, the antigen may be a lipid or carbohydrate. In certain embodiments, the antigen may be a polysaccharide. In certain embodiments, the antigen may be a pneumococcal polysaccharide. In certain embodiments, the polysaccharide may be a Streptococcus pneumoniae polysaccharide. An antigen may be one that generates humoral and / or CTL immune responses.

[0091] As used herein, the term “aryl” refers to a carbocyclic aromatic monocyclic or bicyclic ring system containing about 6 to about 14 carbon atoms. In one embodiment, the aryl group contains about 6 to about 10 carbon atoms. The aryl group may be the same or different and may be substituted with one or more “ring substituents” as defined herein. Non-limiting examples of aryl groups include phenyl and naphthyl. In one embodiment, the aryl group is phenyl. Unless otherwise indicated, the aryl group is unsubstituted.

[0092] As used herein, the term “composition” refers to a formulation containing an active pharmaceutical ingredient or biological component (e.g., Streptococcus pneumoniae polysaccharide carrier protein conjugate and compound) together with one or more additional ingredients. The term “composition” is used interchangeably with “pharmaceutical composition” and “formulation.” A composition may be liquid or solid (e.g., lyophilized). Additional ingredients that may be included as needed include pharmaceutically acceptable excipients, additives, diluents, buffers, sugars, amino acids, chelating agents, surfactants, polyols, bulking agents, stabilizers, cryoprotectants, solubilizers, emulsifiers, salts, adjuvants, isotonic agents, delivery vehicles, and antimicrobial preservatives. A composition is nontoxic to the recipient at the dosage and concentration used.

[0093] As used herein, the term “contains” when used in conjunction with the compositions of the present invention means the inclusion of any other components such as adjuvants and excipients, or the addition of one or more polysaccharide-carrier protein conjugates not specifically enumerated.

[0094] As used herein, the term "consisting of" and variations such as "consist of" when used with polyvalent polysaccharide mixtures or polyvalent polysaccharide carrier protein conjugate mixtures refer to mixtures that have those specific Streptococcus pneumoniae polysaccharides or Streptococcus pneumoniae polysaccharide carrier protein conjugates but do not have other Streptococcus pneumoniae polysaccharides or Streptococcus pneumoniae polysaccharide carrier protein conjugates derived from different serotypes.

[0095] As used herein, the terms "consists essentially of" and variations such as "consist essentially of" or "consisting essentially of" indicate the inclusion of any enumerated element or group of elements, and any other elements of similar or different nature to the enumerated elements, and do not substantially alter the basic or novel properties of the specified dosing regimen, method, or composition.

[0096] As used herein, the terms “de-o-acetylated-15B” or “de-o-acetyl-15B” or “de-O-Ac-15B” refer to de-o-acetylated serotype 15B in which the O-acetyl content is less than 10% per repeating unit. In another embodiment, the O-acetyl content is less than 5% per repeating unit. In another embodiment, the O-acetyl content is less than approximately 1% per repeating unit. In another embodiment, the O-acetyl content is less than 1% per repeating unit. In another embodiment, the O-acetyl content is less than 0.5% per repeating unit. In another embodiment, the O-acetyl content is less than 0.1% per repeating unit. In another embodiment, the O-acetyl content is 0% per repeating unit. The process of de-o-acetylation is known in the art, for example, as described in Rajam et al., Clinical and Vaccine Immunology, 2007, 14(9):1223-1227.

[0097] As used herein, the term “dosage” means the amount of a drug, API (active pharmaceutical ingredient), formulation, composition, pharmaceutical composition, or immunogenic composition that is taken or recommended to be taken at a particular time.

[0098] As used herein, the term “heteroaryl” refers to an aromatic monocyclic or polycyclic ring system containing about 5 to about 14 ring atoms, where 1 to 4 of the ring atoms are independently O, N, or S, and the remaining ring atoms are carbon atoms. In one embodiment, the heteroaryl group has 5 to 10 ring atoms. In another embodiment, the heteroaryl group is monocyclic and has 5 or 6 ring atoms. In another embodiment, the heteroaryl group is bicyclic. The heteroaryl group may be the same or different and may be substituted by one or more “ring system substituents” as defined below herein. The heteroaryl group is linked via ring carbon atoms, and any nitrogen atom of the heteroaryl may be oxidized to the corresponding N-oxide. In one embodiment, the heteroaryl group is a 5-membered heteroaryl. In another embodiment, the heteroaryl group is a 6-membered heteroaryl. In another embodiment, the heteroaryl group includes a 5- to 6-membered heteroaryl group fused to a benzene ring. Unless otherwise indicated, the heteroaryl group is unsubstituted.

[0099] As used herein, the term “heterocycloalkyl” means a saturated or partially unsaturated non-aromatic monocyclic, bicyclic (including spirocyclic), or bridging carbocyclic ring or system containing 3 to about 11 ring atoms, with at least one ring heteroatom selected from N, S, and O, and the remaining ring atoms being carbon atoms. Heterocycloalkyl groups may be bonded via ring carbon or ring nitrogen atoms unless otherwise specified. Heterocycloalkyl rings may be substituted on the ring carbon and / or (one or more) ring nitrogen atoms. In one embodiment, the heterocycloalkyl group is monocyclic and has about 3 to about 7 ring atoms. In another embodiment, the heterocycloalkyl group is monocyclic and has about 4 to about 7 ring atoms. In yet another embodiment, the heterocycloalkyl group is bicyclic and has 7 to 10 ring atoms, 8 to 10 ring atoms, or 9 or 10 ring atoms. In yet another embodiment, the heterocycloalkyl group is monocyclic and has 5 or 6 ring atoms. In one embodiment, the heterocycloalkyl group is monocyclic. In another embodiment, the heterocycloalkyl group is bicyclic. The heterocycloalkyl group can be substituted. In some embodiments, the heterocycloalkyl group has one or two heteroatoms selected from nitrogen, sulfur, and oxygen atoms in the ring. In some embodiments, the heterocycloalkyl group has one heteroatom selected from nitrogen, sulfur, and oxygen atoms in the ring. In some embodiments, the heteroatom is selected from O, S, S(O), S(O)2, and -NH-, -N(alkyl)-. Non-limiting examples include aliphatic groups containing heteroatoms, such as ethers, thioethers, amines, hydroxymethyl, 3-hydroxypropyl, 1,2-dihydroxyethyl, 2-methoxyethyl, 2-aminoethyl, and 2-dimethylaminoethyl.

[0100] As used herein, the terms “immunogenic” or “immunogenic” refer to the ability of an antigen (e.g., streptococcal polysaccharide) to elicit an immune response in a subject. The term “immunogenic composition” refers to the ability of a drug, API, formulation, composition, or pharmaceutical composition to elicit an immune response in a subject.

[0101] As used herein, the term “patients or subjects requiring treatment” includes persons who have been previously exposed to or infected with streptococcal polysaccharide, persons who have been previously vaccinated against streptococcal polysaccharide, and any persons who are susceptible to infection or for whom a reduction in the likelihood of infection is desired, such as immunocompromised individuals, elderly persons, children, adults, or healthy individuals.

[0102] As used herein, the phrase “suitable for the prevention of pneumococcal disease” means that the vaccine or composition is approved by one or more regulatory authorities, such as the U.S. Food and Drug Administration, for the prevention of one or more diseases caused by any serotype of Streptococcus pneumoniae, including but not limited to pneumococcal disease in general, invasive pneumococcal disease (IPD), pneumococcal pneumonia (PP), pneumococcal meningitis, pneumococcal bacteremia, invasive diseases caused by Streptococcus pneumoniae, and otitis media caused by Streptococcus pneumoniae.

[0103] As used herein, the term “multiple doses” refers to a vaccine composition, pharmaceutical composition, or immunogenic composition that requires the administration or injection of more than one dose or components thereof in a clinical regime to induce a sustained immune response, provide protection from disease, or reduce the likelihood of infection by an infectious agent. Those skilled in the art will understand, for example, a method of determining a persistent immune response by measuring antibody titers over a specified period.

[0104] As used herein, “patient” (or “subject” instead) refers to a mammal that is susceptible to infection with Streptococcus pneumoniae polysaccharide. In preferred embodiments, the patient is a human. The patient may be treated prophylactically or therapeutically. Prophylactic treatment provides sufficient protective immunity to reduce the likelihood or severity or effects of pneumococcal infection, e.g., pneumococcal pneumonia. Therapeutic treatment may be performed to reduce the severity of pneumococcal infection or to prevent recurrence or its clinical effects. Prophylactic treatment may be performed using the pneumococcal conjugate composition or vaccine or immunogenic composition of the present invention as described herein. The pneumococcal conjugate composition or vaccine or immunogenic composition of the present invention may be administered to the general population or to persons at high risk of pneumococcal infection, e.g., the elderly, or persons living with or caring for the elderly.

[0105] As used herein, the term "PCV1" refers to a monovalent pneumococcal conjugate vaccine or composition comprising a single Streptococcus pneumoniae polysaccharide carrier protein conjugate containing a capsular polysaccharide derived from a Streptococcus pneumoniae serotype conjugate to a carrier protein. In specific embodiments, the carrier protein is CRM197.

[0106] As used herein, the term "PCV24" refers to a 24-valent pneumococcal conjugate vaccine or composition comprising 23 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a capsular polysaccharide derived from a Streptococcus pneumoniae serotype conjugated to a carrier protein, the serotypes of Streptococcus pneumoniae being at least one of the following: 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 18C, 19A, 19F, 22F, 23B, 24F, 31, 33F, and 35B, as well as serotype 15 being at least one of 15B, 15C, or de-o-acetylated-15B. In certain embodiments, the serotype of serotype 15 is serotype 15C or de-o-acetylated-15B. In other embodiments, the serotype of serotype 15 is serotype de-o-acetylated-15B. In a specific embodiment, one or more carrier proteins in the Streptococcus pneumoniae polysaccharide carrier protein conjugate are CRM197. In a further embodiment, each carrier protein in the Streptococcus pneumoniae polysaccharide carrier protein conjugate is CRM197.

[0107] As used herein, the term "PCV26" refers to a 26-valent pneumococcal conjugate vaccine or composition comprising 25 Streptococcus pneumoniae polysaccharide carrier protein conjugates, each comprising a capsular polysaccharide derived from a Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the serotypes of Streptococcus pneumoniae are at least one of the following: 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 24F, 31, 33F, and 35B, as well as serotype 15: 15B, 15C, or de-O-acetylated-15B. In certain embodiments, the serotype of serotype 15 is serotype 15C or de-O-acetylated-15B. In another embodiment, serotype 15 is serotype de-O-acetylated 15B. In a specific embodiment, one or more carrier proteins in the Streptococcus pneumoniae polysaccharide carrier protein conjugate are CRM197. In a further embodiment, each carrier protein in the Streptococcus pneumoniae polysaccharide carrier protein conjugate is CRM197.

[0108] As used herein, the term “pharmaceutically acceptable” with respect to a carrier, diluent, or excipient of a pharmaceutical composition means that the carrier, diluent, or excipient must be compatible with the other components of the composition and must not be harmful to its recipient.

[0109] As used herein, the term “pharmaceutical composition” refers to a composition containing an active pharmaceutical or biological component together with one or more additional components, for example, a composition in which the active agent is formulated together with one or more pharmaceutically acceptable carriers. As used herein, the terms “pharmaceutical formulation” and “formulation” are used interchangeably with “pharmaceutical composition.” In some embodiments, the active agent is present in a unit dose appropriate for administration in a therapeutic regime that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to the relevant population. The pharmaceutical composition or formulation may be liquid or solid (e.g., lyophilized). Additional components that may be included as needed include pharmaceutically acceptable excipients, additives, diluents, buffers, sugars, amino acids, chelating agents, surfactants, polyols, bulking agents, stabilizers, cryoprotectants, solubilizers, emulsifiers, salts, adjuvants, isotonic agents, delivery vehicles, and antimicrobial preservatives. The pharmaceutical composition or formulation is nontoxic to the recipient at the dose and concentration used. In some embodiments, pharmaceutical compositions can be specifically formulated for administration in solid or liquid form, including: oral administration, e.g., oral tablets (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeting buccal, sublingual, and systemic absorption, boluses, powders, granules, and pastes for application to the tongue; parenteral administration, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection, e.g., sterile solutions or suspensions, or sustained-release formulations; topical application, e.g., as creams, ointments, or controlled-release patches or sprays applied to the skin, lungs, or oral cavity; intravaginal or rectal, e.g., as pessaries, creams, or foams; sublingual; ocular; transdermal; or transnasal, transpulmonary, and to other mucosal surfaces. In some embodiments, the term "formulation" refers to a single dose of vaccine that can be contained in any volume suitable for injection.

[0110] As used herein, the terms “pneumococcal conjugate” or “pneumococcal polysaccharide carrier protein conjugate” refer to the pneumococcal polysaccharide carrier protein conjugate.

[0111] As used herein, the term “pneumococcal conjugate vaccine” (or “PCV”) refers to a pharmaceutical formulation or composition comprising one or more pneumococcal polysaccharide carrier protein conjugates that provide active immunity to a disease or pathological condition caused by one or more serotypes of Streptococcus pneumoniae.

[0112] As used herein, the term “ring substituent” refers to a substituent bonded to an aromatic or non-aromatic ring system that replaces, for example, available hydrogens on the ring system. Ring substituents may be the same or different, and each may be independently selected. Examples of ring substituents include alkyl, alkenyl, alkynyl, aryl, heteroaryl, -OH, hydroxyalkyl, haloalkyl, -O-alkyl, -O-haloalkyl, -alkylene-O-alkyl, -O-aryl, -O-alkylene-aryl, acyl, -C(O)-aryl, halo, -NO2, -CN, -SF5, -C(O)OH, -C(O)O-alkyl, -C(O)O-aryl, -C(O)O-alkylene-aryl, -S(O)-alkyl, -S(O)2-aryl Examples of cyclic substituents include chlorine, -S(O)-aryl, -S(O)2-aryl, -S(O)-heteroaryl, -S(O)2-heteroaryl, -S-alkyl, -S-aryl, -S-heteroaryl, -S-alkylene-aryl, -S-alkylene-heteroaryl, -S(O)2-alkylene-aryl, -S(O)2-alkylene-heteroaryl, cycloalkyl, heterocycloalkyl, -OC(O)-alkyl, -OC(O)-aryl, and -OC(O)-cycloalkyl. Further examples of cyclic substituents include (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0113] As used herein, the term “single dose” refers to a vaccine composition that requires only one administration or injection in a clinical regime to induce a sustained immune response and provide protection from a disease (e.g., pneumococcal disease). Those skilled in the art will understand, for example, how to determine a persistent immune response by measuring antibody titers over a specified period.

[0114] As used herein, the terms “substituent,” “any substituent,” or “may be substituted” refer, for example, to alkyl groups, alkenyl groups, or substituents bonded to alkynyl groups that replace available hydrogens on the group. Substituents may be the same or different, and each is independently selected. Examples of substituents include alkyl, alkenyl, alkynyl, aryl, heteroaryl, -OH, hydroxyalkyl, haloalkyl, -O-alkyl, -O-haloalkyl, -alkylene-O-alkyl, -O-aryl, -O-alkylene-aryl, acyl, -C(O)-aryl, halo, -NO2, -CN, -SF5, -C(O)OH, -C(O)O-alkyl, -C(O)O-aryl, -C(O)O-alkylene-aryl, -S(O)-alkyl, -S(O)2-alkyl Examples of substituents include -S(O)-aryl, -S(O)2-aryl, -S(O)-heteroaryl, -S(O)2-heteroaryl, -S-alkyl, -S-aryl, -S-heteroaryl, -S-alkylene-aryl, -S-alkylene-heteroaryl, -S(O)2-alkylene-aryl, -S(O)2-alkylene-heteroaryl, cycloalkyl, heterocycloalkyl, -OC(O)-alkyl, -OC(O)-aryl, and -OC(O)-cycloalkyl. Further examples of substituents include (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0115] As used herein, “SNE” or “stable nanoemulsion” refers to a composition comprising a compound of formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof. In some embodiments, the SNE comprises a compound of formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, sorbitan trioleate (SPAN-85), polysorbate-20 (PS-20) or polysorbate-80 (PS-80), and squalene, and the composition is in the form of a nanoemulsion. In some embodiments, SNE comprises a compound of formula I, Ia, II, IIa, III, IIIa, IV, or IVa, or a pharmaceutically acceptable salt thereof, SPAN-85, PS-20, and squalene. In some embodiments, SNE comprises 0.001 mg / mL to 10 mg / mL of SPAN-85, 0.001 mg / mL to 10 mg / mL of PS-20, and 0.01 mg / mL to 100 mg / mL of squalene. In further embodiments, SNE comprises 0.05 mg / mL to 5 mg / mL of SPAN-85, 0.05 mg / mL to 5 mg / mL of PS-20, and 0.05 mg / mL to 50 mg / mL of squalene. In further embodiments, the SNE comprises 0.1 mg / mL to 1 mg / mL of SPAN-85, 0.1 mg / mL to 1 mg / mL of PS-20, and 1 mg / mL to 10 mg / mL of squalene. As used herein, the SNE is used interchangeably with "squalene emulsion".

[0116] Nanoemulsions, also known as nanometer-sized emulsions, are fine oil-in-water (o / w) dispersions of two immiscible fluids. Nanoemulsions are submicron-sized colloidal particle systems that act as carriers for drug molecules. Their sizes vary from 10 to 1,000 nm. These carriers are solid spheres with amorphous and lipophilic surfaces.

[0117] As used herein, the term “therapeutic dose” means the amount of an active component (antigen) sufficient to produce a desired therapeutic effect in a human or animal, for example, the amount necessary to induce an immune response, treat, cure, prevent or inhibit the onset and progression of a disease or its symptoms, and / or the amount necessary to improve symptoms or cause disease regression. The therapeutic dose may vary depending on the structure and potency of the active component and the intended mode of administration. Those skilled in the art can readily determine the therapeutic dose of a given active component in a vaccine.

[0118] As used herein, the term “valence” refers to the presence of a specified number of polysaccharides or polysaccharide-carrier protein conjugates in a composition.

[0119] As used herein, the terms “vaccine” or “vaccine composition” refer to a biological product used to stimulate antibody production and provide immunity against an infectious disease.

[0120] As used herein, the term “carbon or nitrogen-linked spacer” refers to any chemistry that links carbon (-C) or nitrogen (-N) to the benzyl group of the formulas of the present invention (see the linkage of the variable “A” in formulas I and Ia). Examples of carbon or nitrogen-linked spacers are C1-C6 alkyl, heterocycloalkyl, aryl, and heteroaryl, wherein the alkyl, heterocycloalkyl, aryl, and heteroaryl may be substituted.

[0121] Unless otherwise specified, when describing a linker throughout this disclosure, the first available bond on the linker group is attached to the left portion of the compound adjacent to the linker group, and the terminal available bond on the linker group is attached to the right portion of the compound adjacent to the linker group. For example, if the linker (L) is -CH2-CF2-, the definition of L includes only A-CH2-CF2-B and does not include A-CF2-CH2-B.

[0122] As used herein, the term “functional group” refers to any chemical property that links a carbon or nitrogen-linked spacer to a lipid group. Examples of functional groups are: [ka] The equation is such that n is 0, 1, 2, 3, 4, or 5.

[0123] Further examples of functional groups are: [ka] That is the case.

[0124] As used herein, the term “lipid” refers to any chemical property that is insoluble in water but soluble in organic solvents. Examples of lipids are represented by variable D (see formulas I and Ia), (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) Selected from alkynyls, the alkyl, alkenyl, and alkynyl may be substituted.

[0125] Further examples of lipids are, [ka] And, Any carbon atom on the lipid chain may be substituted. [ka] This is cis or trans stereochemistry, X is O, C(R) 2 , or NR, R is independently selected from H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, OH, O(C1-C4)alkyl, O(C1-C4)alkenyl, O(C1-C4)alkynyl, chlorine, and fluorine.

[0126] Further examples of lipids are represented by variable D (see formulas I and Ia), where D is

Chemical structure

[0127] Compounds of the invention having one or more asymmetric centers may exist, unless otherwise specified, as mixtures of stereoisomers or as substantially pure individual diastereomers or enantiomers. The invention encompasses all stereoisomeric forms of the compounds of formulas I, Ia, II, IIa, III, IIIa, IV, and IVa. Unless a specific stereochemistry is shown, the invention is meant to include all such isomers of these compounds. The asymmetric centers present in the compounds of formulas I, Ia, II, IIa, III, IIIa, IV, and IVa can each independently have the (R) configuration or the (S) configuration. When a bond to a chiral carbon is shown as a straight line in the structural formula of the invention, it is understood that both the (R) and (S) configurations of the chiral carbon, and thus both the enantiomers and mixtures thereof, are included in the formula. Similarly, when a compound name is listed without chiral designation for a chiral carbon, it is understood that both the (R) and (S) configurations of the chiral carbon, and thus the individual enantiomers, diastereomers, and mixtures thereof, are included in that name. The production of specific stereoisomers or mixtures thereof can be identified in the examples in which such stereoisomers or mixtures were obtained, but this in no way limits the scope of the invention to the inclusion of all stereoisomers and mixtures thereof.

[0128] If any component or any variable (e.g., n, Rb, etc.) appears two or more times in formulas I, Ia, II, IIa, III, IIIa, IV, and IVa, its definition at each appearance is independent of its definition at all other appearances. Furthermore, combinations of substituents and / or variables are only permitted if such combinations result in a stable compound.

[0129] A “stable” compound is one that can be prepared and isolated and remains essentially unchanged for a period of time sufficient to allow the compound to be used for the purposes described herein (e.g., as an adjuvant in an immunogenic composition administered to a subject), while retaining its structure and properties. The compounds of the present invention are limited to stable compounds encompassed by formulas I, Ia, II, IIa, III, IIIa, IV, and IVa.

[0130] The dashed lines used in this specification [ka] The lines indicate the bonding points with the rest of the compound. Lines drawn on the ring system, for example: [ka] This indicates that the bond can be attached to any of the replaceable ring atoms.

[0131] compound The compounds described herein are useful in adjuvant formulations of the present invention that are useful for boosting the immunological response of the pneumococcal composition or pneumococcal conjugate composition or pneumococcal vaccine (PV) or pneumococcal conjugate vaccine (PCV) described herein.

[0132] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I: [ka] (wherein, R a is selected from H, -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, and -NR’R’’, and the (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, and -O(C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine, R a’ is selected from H, -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, and -NR’R’’, and the (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, and -O(C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine, R a’’ is selected from H, -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, and -NR’R’’, and the (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, and -O(C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine, R' and R'' are independently selected from H, (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl, and the (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine, or R' and R'' together with the nitrogen to which they are bonded to form a (C3-C6) heterocycloalkyl, and the (C3-C6) heterocycloalkyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. R b Each occurrence is independently selected from H, -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, or NR'R'', and the (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, and -O(C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. A is (C1-C6) alkyl, heterocycloalkyl, heterocycloalkyl-C(O)-R z -, (C1-C4)alkyl-N(R z )-R z-A carbon or nitrogen bond spacer selected from aryl and heteroaryl groups, wherein the (C1-C6)alkyl, heterocycloalkyl, aryl, and heteroaryl groups are independently selected from the group consisting of -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, and NR'R''. The (C1-C6) alkyl, (C1-C6) alkenyl, (C1-C6) alkynyl, -O(C1-C6) alkyl, -O(C1-C6) alkenyl, and -O(C1-C6) alkynyl may be substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4) alkyl, -O(C1-C4) alkenyl, -O(C1-C4) alkynyl, chlorine, and fluorine. R z Each occurrence is independently H or (C1-C6) alkyl. B is [ka] A functional group selected from, D is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 )A lipid selected from alkynyl, and the above (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 The alkynyl group may be substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine, or D is [ka] And, Each occurrence of Z is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20) independently selected from alkynyl, and the (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 The alkynyl group may be substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. m is 0, 1, 2, 3, 4, or 5. n is 0, 1, 2, 3, 4, or 5.

[0133] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where m is 0, 1, 2, 3, 4, or 5.

[0134] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where m is 0, 1, 2, 3, or 4.

[0135] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where m is 0, 1, 2, or 3.

[0136] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where m is 0, 1, or 2.

[0137] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where m is 2.

[0138] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where m is 1.

[0139] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where m is 0.

[0140] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where n is 0, 1, 2, 3, 4, or 5.

[0141] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where n is 0, 1, 2, 3, or 4.

[0142] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where n is 0, 1, 2, or 3.

[0143] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where n is 0, 1, or 2.

[0144] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where n is 2.

[0145] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where n is 1.

[0146] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where n is 0.

[0147] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where R' and R'' are independently H, (C1-C6)alkyl, (C1-C6)alkenyl, or (C1-C6)alkynyl, and the (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0148] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where R' and R'' are independently H, (C1-C6)alkyl, (C1-C6)alkenyl, or (C1-C6)alkynyl, and the (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from -OH and -O(CH3).

[0149] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where R' and R'' are independently H, (C1-C4)alkyl, (C1-C4)alkenyl, or (C1-C4)alkynyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, and (C1-C4)alkynyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0150] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where R' and R'' are independently H and (C1-C4)alkyl, and the (C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0151] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula I, where R' and R'' are independently H and (C1-C4)alkyl.

[0152] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, wherein R' and R'' together with the nitrogen to which they are bound form a (C3-C6) heterocycloalkyl, and the (C3-C6) heterocycloalkyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0153] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a is H, -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, or NR'R'', wherein the (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, and -O(C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0154] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R ais H, -OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, fluorine, or NR'R'', wherein the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, and -O(C1-C4)alkynyl may be substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0155] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a is H, -OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, fluorine, or NR'R'', wherein the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, and -O(C1-C4)alkynyl may be substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0156] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a is H, -OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, fluorine, or NR'R''.

[0157] In some embodiments, the compounds of the present invention are represented by the structure shown in formula I, where R a It is NR'R''.

[0158] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a It is NR'R''.

[0159] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is pentyl.

[0160] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is butyl.

[0161] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is propyl.

[0162] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is ethyl.

[0163] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is methyl.

[0164] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is H.

[0165] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a’ is H, -OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, fluorine, or NR'R'', wherein the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, and -O(C1-C4)alkynyl may be substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0166] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a’ is H, -OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, fluorine, or NR'R'', wherein the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, and -O(C1-C4)alkynyl may be substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0167] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a’is H, -OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, fluorine, or NR'R''.

[0168] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a’ It is NR'R''.

[0169] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is pentyl.

[0170] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is butyl.

[0171] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is propyl.

[0172] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is ethyl.

[0173] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is methyl.

[0174] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is H.

[0175] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a’’ is H, -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, or NR'R'', wherein the (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, and -O(C1-C6)alkynyl may be substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0176] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a’’ is H, -OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, fluorine, or NR'R'', wherein the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, and -O(C1-C4)alkynyl may be substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0177] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a’’is H, -OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, fluorine, or NR'R''.

[0178] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a’’ It is NR'R''.

[0179] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is pentyl.

[0180] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is butyl.

[0181] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is propyl.

[0182] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is ethyl.

[0183] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is methyl.

[0184] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R' is H and R'' is H.

[0185] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R a’’ H is H.

[0186] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R b is H, -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, or NR'R'', wherein the (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, and -O(C1-C6)alkynyl may be substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0187] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R b is H, -OH, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, fluorine, or NR'R'', wherein the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, and -O(C1-C4)alkynyl may be substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0188] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R b These are independently H, -OH, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, or NR'R''.

[0189] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R b These are independently H, -OH, (C1-C4)alkyl, -O(C1-C4)alkyl, chlorine, fluorine, or NH2.

[0190] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where R b These are independently H and O(CH3).

[0191] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where A is (C1-C6)alkyl or heterocycloalkyl, and the alkyl or heterocycloalkyl is -OH, oxo, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, -O(C1-C6)alkynyl, chlorine, fluorine, and NR'. The (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, and -O(C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0192] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where A is (C1-C4)alkyl or heterocycloalkyl, and the alkyl or heterocycloalkyl is -OH, oxo, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, fluorine, and NR'. The (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, and -O(C1-C4)alkynyl may be substituted with 1 to 6 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0193] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where A is heterocycloalkyl-C(O)-R z - or (C1-C4)alkyl-N(R z ) 2 - and R z Each occurrence is independently either H or (C1-C6) alkyl.

[0194] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, wherein A is [ka] (In the formula, R z These are independently selected from H, (C1-C6)alkyl, (C1-C6)alkenyl, (C1-C6)alkynyl, -O(C1-C6)alkyl, -O(C1-C6)alkenyl, and -O(C1-C6)alkynyl. X is either CH or N, R d This is independently selected from H, -OH, oxo, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. n is 0, 1, 2, 3, or 4. p is selected from 0, 1, 2, 3, or 4.

[0195] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, wherein A is [ka] (In the formula, R z These are independently selected from H, (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl. X is either CH or N, R d This is independently selected from -OH, (C1-C4)alkyl, -O(C1-C4)alkyl, chlorine, and fluorine. n is 0, 1, 2, or 3. p is selected from 0, 1, or 2.

[0196] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, wherein A is [ka] (In the formula, R z These are independently H or (C1-C6) alkyl, X is either CH or N, R d This is independently selected from -OH, (C1-C4)alkyl, -O(C1-C4)alkyl, chlorine, and fluorine. n is 0, 1, 2, or 3. p is selected from 0, 1, or 2.

[0197] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, wherein A is [ka] (In the formula, R z These are independently H or (C1-C6) alkyl, R d This is independently selected from -OH, (C1-C4)alkyl, -O(C1-C4)alkyl, chlorine, and fluorine. n is 0, 1, 2, or 3. p is selected from 0, 1, or 2.

[0198] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, wherein A is [ka] (In the formula, R z This is independently selected from H or (C1-C6) alkyl.

[0199] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, wherein B is [ka] Selected from.

[0200] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, wherein B is [ka] That is the case.

[0201] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where D is (C6-C 20 ) alkyl, (C6-C 20 ) Alkenyl, or (C6-C 20 ) is an alkynyl, and the above (C6-C 20 ) alkyl, (C6-C 20 ) Alkenyl, or (C6-C 20 The alkynyl group may be substituted with 1 to 6 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine, or D is [ka] (In the formula, Z is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) Selected from alkynyl, and the above (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 The alkynyl group may be substituted with 1 to 6 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0202] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, where D is (C6-C 20 ) alkyl, (C6-C 20 ) Alkenyl, or (C6-C 20 ) is alkinyl, or D is [ka] (In the formula, Z is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 (Selected from Alkinil).

[0203] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, wherein D is [ka] (In the formula, any carbon on the lipid chain may be substituted with -OH, -O(CH3), chlorine, or fluorine.) [ka] This is cis or trans stereochemistry, X is O, C(R) 2 , or NR, R is selected independently from H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine.

[0204] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula I, wherein D is [ka] (In the formula, any carbon on the lipid chain may be substituted with -OH, -O(CH3), chlorine, or fluorine.) [ka] This is cis or trans stereochemistry, X 1 -O-, -C(R) 2 -, or -NR- Each R occurrence is independently selected from H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. n is 0, 1, 2, 3, 4, or 5. q is 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9. s is 0, 1, 2, 3, 4, 5, 6, 7, or 8. t is selected from 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18.

[0205] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia: [ka] (In the formula, R' and R'' are independently selected from H, (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl, and the (C1-C6)alkyl, (C1-C6)alkenyl, and (C1-C6)alkynyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine, or R' and R'' together with the nitrogen to which they are bonded to form a (C3-C6) heterocycloalkyl, and the (C3-C6) heterocycloalkyl may be substituted with 1 to 4 substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. R b Each occurrence is -O(C1-C4)alkyl, and the -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from the group consisting of -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. A is [ka] Selected from, Rz Each occurrence is independently H or (C1-C6) alkyl. R d Each occurrence is independently selected from -OH, (C1-C4)alkyl, -O(C1-C4)alkyl, chlorine, and fluorine. B is [ka] And, D is [ka] A lipid chain selected from the following, where any carbon on the lipid chain may be substituted with -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, or fluorine. [ka] It is cis or trans stereochemistry, X 1 -O-, -C(R) 2 -, or -NR- Each R occurrence is independently selected from H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. m is 0, 1, or 2. n is 0, 1, 2, or 3. p is 0, 1, or 2. q is 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9. s is 1, 2, 3, 4, 5, 6, 7, or 8. t is 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18).

[0206] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, where m is 2.

[0207] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, where m is 1.

[0208] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, where m is 0.

[0209] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, wherein R' and R'' are independently selected from H and (C1-C6) alkyl groups, and the alkyl group may be substituted with 1 to 4 -OH and / or -O(CH3) groups.

[0210] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, wherein R' and R'' are independently selected from H and (C1-C4) alkyl groups, and the alkyl group may be substituted with 1 to 4 substituents independently selected from -OH and -O(CH3).

[0211] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, where R' is H and R'' is pentyl.

[0212] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, where R' is H and R'' is butyl.

[0213] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, where R' is H and R'' is propyl.

[0214] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, where R' is H and R'' is ethyl.

[0215] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, where R' is H and R'' is methyl.

[0216] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, where R' is H and R'' is H.

[0217] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, wherein R a’’ is H or (C1-C4) alkyl, and the alkyl may be substituted with one or two independently selected -OH or -O(CH3).

[0218] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, wherein R a’’ is H or (C1-C4) alkyl.

[0219] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, wherein R a’’ H is H.

[0220] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, wherein R b is an -O(C1-C4)alkyl group, and the -O(C1-C4)alkyl group may be substituted with one or two independently selected -OH or -O(CH3) groups.

[0221] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, wherein R b It is -O(CH3).

[0222] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula Ia, wherein A is [ka] Selected from, R z These are independently H or (C1-C6) alkyl.

[0223] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula II: [ka] (In the formula, R 1 is a (C1-C6)alkyl group, and the (C1-C6)alkyl group may be substituted with 1 to 4 substituents independently selected from -OH and -O(CH3). R 2 is H, methyl or -O(CH3), R 3Each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH and -O(CH3). R 4 Each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH and -O(CH3). R 5 teeth, [ka] And, R 6 (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) Selected from alkynyl, and the above (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 The alkynyl group may be substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. Each occurrence of n is 4.

[0224] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula II, where each occurrence of n is independently 0, 1, 2, or 3.

[0225] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula II, where each occurrence of n is independently 0, 1, or 2.

[0226] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula II, where each occurrence of n is independently 0 or 1.

[0227] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein n is absent.

[0228] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 1 is ethyl, butyl (branched or linear), or pentyl (branched or linear), and the ethyl, butyl, or pentyl may be substituted with 1 to 4 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0229] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 1 The molecule is butyl (branched or linear), and the butyl may be substituted with 1 to 4 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0230] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 1 is ethyl, butyl (branched or linear), or pentyl (branched or linear), and the ethyl, butyl, or pentyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0231] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 1 The molecule is butyl (branched or straight), and the butyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0232] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 2 It is either H or methyl.

[0233] In some embodiments, the compounds of the present invention are represented by the structure shown in formula II, where R 2 H is H.

[0234] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 3 Each occurrence is independently a (C1-C4)alkyl, (C1-C4)alkenyl, or -O(C1-C4)alkyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, or -O(C1-C4)alkyl may be substituted with 1 to 4 substituents independently selected from -OH or -O(CH3).

[0235] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 3 Each occurrence is independently (C1-C4)alkyl or -O(C1-C4)alkyl, and the (C1-C4)alkyl or -O(C1-C4)alkyl may be substituted with 1 to 4 substituents independently selected from -OH or -O(CH3).

[0236] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R3 Each occurrence is independently (C1-C4)alkyl or -O(C1-C4)alkyl, and the (C1-C4)alkyl or -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH or -O(CH3).

[0237] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 3 Each occurrence is independently either methyl or -O(CH3).

[0238] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 3 Each occurrence of this expression is -O(CH3).

[0239] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 4 Each occurrence is independently a (C1-C4)alkyl or -O(C1-C4)alkyl, and the (C1-C4)alkyl or -O(C1-C4)alkyl may be substituted with 1 to 4 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0240] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 4 Each occurrence is independently (C1-C4)alkyl or -O(C1-C4)alkyl.

[0241] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 (C6-C 20 )alkyl and (C6-C 20 ) Selected from alkenyls, and the above (C6-C20 )alkyl and (C6-C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0242] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 (C8-C 20 )alkyl and (C8-C 20 ) Selected from alkenyls, and the above (C8-C 20 )alkyl and (C8-C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0243] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 is, (C 10 -C 20 )alkyl and (C 10 -C 20 ) Selected from alkenyls, and the (C 10 -C 20 )alkyl and (C 10 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0244] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 is, (C 12 -C 20 )alkyl and (C 12 -C 20 ) Selected from alkenyls, and the (C 12 -C 20 )alkyl and (C 12 -C 20The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0245] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 is, (C 14 -C 20 )alkyl and (C 14 -C 20 ) Selected from alkenyls, and the (C 14 -C 20 )alkyl and (C 14 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0246] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 is, (C 16 -C 20 )alkyl and (C 16 -C 20 ) Selected from alkenyls, and the (C 16 -C 20 )alkyl and (C 16 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0247] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 (C6-C 20 )alkyl and (C6-C 20 ) Selected from Alkenil.

[0248] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 (C8-C20 )alkyl and (C8-C 20 ) Selected from Alkenil.

[0249] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 is (C 10 -C 20 )alkyl and (C 10 -C 20 ) Selected from Alkenil.

[0250] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 is, (C 12 -C 20 )alkyl and (C 12 -C 20 ) Selected from Alkenil.

[0251] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 is (C 14 -C 20 )alkyl and (C 14 -C 20 ) Selected from Alkenil.

[0252] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula II, wherein R 6 is, (C 16 -C 20 )alkyl and (C 16 -C 20 ) Selected from Alkenil.

[0253] The present invention further provides compositions comprising a compound having the structure described in formula IIa: [ka] (In the formula, R 1 is butyl, and the butyl may be substituted with one or two -OH groups. R 3 Each occurrence is independently either H or -O(CH3), R 5 teeth, [ka] And, R 6 is, (C 10 -C 20 ) alkyl, (C 10 -C 20 ) Alkenyl, and (C 10 -C 20 (Selected from Alkinil).

[0254] In some embodiments, the compounds of the present invention are represented by the structure shown in formula IIa, where R 6 is, (C 10 -C 20 )alkyl and (C 10 -C 20 ) Selected from alkenyls, and the (C 10 -C 20 )alkyl and (C 10 -C 20 The alkenyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0255] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIa, where R 6 is, (C 12 -C 20 )alkyl and (C 12 -C 20 ) Selected from alkenyls, and the (C 12 -C 20 )alkyl and (C 12 -C 20The alkenyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0256] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIa, where R 6 is, (C 14 -C 20 )alkyl and (C 14 -C 20 ) Selected from alkenyls, and the (C 14 -C 20 )alkyl and (C 14 -C 20 The alkenyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0257] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIa, where R 6 is, (C 16 -C 20 )alkyl and (C 16 -C 20 ) Selected from alkenyls, and the (C 16 -C 20 )alkyl and (C 16 -C 20 The alkenyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0258] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIa, where R 6 is, (C 10 -C 20 )alkyl and (C 10 -C 20 ) Selected from Alkenil.

[0259] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIa, where R 6 is, (C 12 -C 20 )alkyl and (C 12 -C 20 ) Selected from Alkenil.

[0260] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIa, where R 6 is, (C 14 -C 20 )alkyl and (C 14 -C 20 ) Selected from Alkenil.

[0261] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIa, where R 6 is, (C 16 -C 20 )alkyl and (C 16 -C 20 ) Selected from Alkenil.

[0262] The present invention provides a composition comprising a compound having the structure described in Formula III: [ka] (In the formula, R 1 is a (C1-C6)alkyl group, and the (C1-C6)alkyl group may be substituted with 1 to 4 substituents independently selected from -OH and -O(CH3). R 2 is H, methyl or -O(CH3), R 3Each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH and -O(CH3). R 4 (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) Selected from alkynyl, and the above (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 The alkynyl group may be substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, and fluorine. n is 4).

[0263] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula III, where n is 0, 1, 2, or 3.

[0264] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula III, where n is 0, 1, or 2.

[0265] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula III, where n is 0 or 1.

[0266] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula III, wherein n is absent.

[0267] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 1 is ethyl, butyl (branched or linear), or pentyl (branched or linear), and the ethyl, butyl, or pentyl may be substituted with 1 to 4 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0268] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 1 The molecule is butyl (branched or linear), and the butyl may be substituted with 1 to 4 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0269] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 1 is ethyl, butyl (branched or linear), or pentyl (branched or linear), and the ethyl, butyl, or pentyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0270] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 1 The molecule is butyl (branched or straight), and the butyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0271] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 2 It is either H or methyl.

[0272] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 2 H is H.

[0273] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 3 Each occurrence is independently a (C1-C4)alkyl, (C1-C4)alkenyl, or -O(C1-C4)alkyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, or -O(C1-C4)alkyl may be substituted with 1 to 4 substituents independently selected from -OH or -O(CH3).

[0274] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 3 Each occurrence is independently (C1-C4)alkyl or -O(C1-C4)alkyl, and the (C1-C4)alkyl or -O(C1-C4)alkyl may be substituted with 1 to 4 substituents independently selected from -OH or -O(CH3).

[0275] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 3 Each occurrence is independently (C1-C4)alkyl or -O(C1-C4)alkyl, and the (C1-C4)alkyl or -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH or -O(CH3).

[0276] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 3 Each occurrence is independently either methyl or -O(CH3).

[0277] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 3 Each occurrence of this expression is -O(CH3).

[0278] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 (C6-C 20 )alkyl and (C6-C 20 ) Selected from alkenyls, and the above (C6-C 20 )alkyl and (C6-C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0279] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 (C8-C 20 )alkyl and (C8-C 20 ) Selected from alkenyls, and the above (C8-C 20 )alkyl and (C8-C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0280] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 is, (C 10 -C 20 )alkyl and (C 10 -C 20 ) Selected from alkenyls, and the (C 10 -C 20 )alkyl and (C 10 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0281] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 is, (C 12 -C 20 )alkyl and (C 12 -C 20 ) Selected from alkenyls, and the (C 12 -C 20 )alkyl and (C 12 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0282] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 is, (C 14 -C 20 )alkyl and (C 14 -C 20 ) Selected from alkenyls, and the (C 14 -C 20 )alkyl and (C 14 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0283] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 is, (C 16 -C 20 )alkyl and (C 16 -C 20 ) Selected from alkenyls, and the (C 16 -C 20 )alkyl and (C 16 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0284] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 (C6-C 20 )alkyl and (C6-C 20 ) Selected from Alkenil.

[0285] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 (C8-C 20 )alkyl and (C8-C 20 ) Selected from Alkenil.

[0286] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 is, (C 10 -C 20 )alkyl and (C 10 -C 20 ) Selected from Alkenil.

[0287] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 is, (C 12 -C 20 )alkyl and (C 12 -C 20 ) Selected from Alkenil.

[0288] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 is, (C 14 -C 20 )alkyl and (C 14 -C 20 ) Selected from Alkenil.

[0289] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula III, where R 4 is, (C 16 -C 20 )alkyl and (C 16 -C 20 ) Selected from Alkenil.

[0290] The present invention also provides compositions comprising compounds having the structure described in formula IIIa: [ka] (In the formula, R 1 is butyl, and the butyl may be substituted with one or two -OH groups. R 3 Each occurrence is independently either H or -O(CH3), R 4 is, (C 10 -C 20 ) alkyl, (C 10 -C 20 ) Alkenyl, and (C 10 -C 20 (Selected from Alkinil).

[0291] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 (C6-C 20 )alkyl and (C6-C 20 ) Selected from alkenyls, and the above (C6-C 20 )alkyl and (C6-C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0292] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 (C8-C 20)alkyl and (C8-C 20 ) Selected from alkenyls, and the above (C8-C 20 )alkyl and (C8-C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0293] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 is, (C 10 -C 20 )alkyl and (C 10 -C 20 ) Selected from alkenyls, and the (C 10 -C 20 )alkyl and (C 10 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0294] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 is, (C 12 -C 20 )alkyl and (C 12 -C 20 ) Selected from alkenyls, and the (C 12 -C 20 )alkyl and (C 12 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0295] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 is, (C 14 -C 20 )alkyl and (C 14 -C 20 ) Selected from alkenyls, and the (C14 -C 20 )alkyl and (C 14 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0296] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 is, (C 16 -C 20 )alkyl and (C 16 -C 20 ) Selected from alkenyls, and the (C 16 -C 20 )alkyl and (C 16 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0297] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 (C6-C 20 )alkyl and (C6-C 20 ) Selected from Alkenil.

[0298] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 (C8-C 20 )alkyl and (C8-C 20 ) Selected from Alkenil.

[0299] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 is, (C 10 -C 20 )alkyl and (C 10 -C 20) Selected from Alkenil.

[0300] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 is, (C 12 -C 20 )alkyl and (C 12 -C 20 ) Selected from Alkenil.

[0301] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 is, (C 14 -C 20 )alkyl and (C 14 -C 20 ) Selected from Alkenil.

[0302] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IIIa, where R 4 is, (C 16 -C 20 )alkyl and (C 16 -C 20 ) Selected from Alkenil.

[0303] The present invention provides a composition comprising a compound having the structure described in Formula IV: [ka] (In the formula, R 1 is a (C1-C6)alkyl group, and the (C1-C6)alkyl group may be substituted with 1 to 4 substituents independently selected from -OH and -O(CH3). R 2 is H, methyl or -O(CH3), R 3Each occurrence is independently H, (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, or -O(C1-C4)alkyl, and the (C1-C4)alkyl, (C1-C4)alkenyl, (C1-C4)alkynyl, and -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH and -O(CH3). R 4 Each occurrence is (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 ) independently selected from alkynyl, and the (C6-C 20 ) alkyl, (C6-C 20 ) alkenyl, and (C6-C 20 The alkynyl group may be substituted with 1 to 6 substituents independently selected from -OH, -O(C1-C4)alkyl, -O(C1-C4)alkenyl, -O(C1-C4)alkynyl, chlorine, or fluorine. n is 4).

[0304] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where n is 0, 1, 2, or 3.

[0305] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where n is 0, 1, or 2.

[0306] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where n is 0 or 1.

[0307] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, wherein n is absent.

[0308] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 1 is ethyl, butyl (branched or linear), or pentyl (branched or linear), and the ethyl, butyl, or pentyl may be substituted with 1 to 4 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0309] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 1 is butyl (branched or linear), and the butyl may be substituted with 1 to 4 substituents independently selected from -OH, -O(CH3), chlorine, and fluorine.

[0310] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 1 is ethyl, butyl (branched or linear), or pentyl (branched or linear), and the ethyl, butyl, or pentyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0311] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 1 is butyl (branched or linear), and the butyl may be substituted with one or two substituents independently selected from -OH and -O(CH3).

[0312] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 2 It is either H or methyl.

[0313] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 2 H is H.

[0314] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 3 The (C1-C4)alkyl, (C1-C4)alkenyl, or -O(C1-C4)alkyl group is independently a (C1-C4)alkyl group, and the (C1-C4)alkyl group, (C1-C4)alkenyl, or -O(C1-C4)alkyl group may be substituted with 1 to 4 substituents independently selected from -OH or -O(CH3).

[0315] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula IV, R 3 Each occurrence is independently (C1-C4)alkyl or -O(C1-C4)alkyl, and the (C1-C4)alkyl or -O(C1-C4)alkyl may be substituted with 1 to 4 substituents independently selected from -OH or -O(CH3).

[0316] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in Formula IV, R 3 Each occurrence is independently (C1-C4)alkyl or -O(C1-C4)alkyl, and the (C1-C4)alkyl or -O(C1-C4)alkyl may be substituted with one or two substituents independently selected from -OH or -O(CH3).

[0317] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 3 Each occurrence is independently either methyl or -O(CH3).

[0318] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 3 Each occurrence of this expression is -O(CH3).

[0319] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C6-C 20 )alkyl and (C6-C 20 ) independently selected from alkenyls, and the (C6-C 20 )alkyl and (C6-C 20 The alkenyl may be substituted with 1 to 6 independently selected -OH or -O(CH3) groups.

[0320] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C8-C 20 )alkyl and (C8-C 20 ) independently selected from alkenyls, and the (C8-C 20 )alkyl and (C8-C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0321] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C 10 -C 20 )alkyl and (C 10 -C 20 ) independently selected from alkenyl, and the (C 10 -C 20 )alkyl and (C 10 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0322] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C 12 -C 20 )alkyl and (C 12 -C 20 ) independently selected from alkenyl, and the (C 12 -C 20 )alkyl and (C 12 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0323] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C 14 -C 20 )alkyl and (C 14 -C 20 ) independently selected from alkenyl, and the (C 14 -C 20 )alkyl and (C 14 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0324] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C 16 -C 20 )alkyl and (C 16 -C 20 ) independently selected from alkenyl, and the (C 16 -C 20 )alkyl and (C 16 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0325] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C6-C 20 )alkyl and (C6-C 20 ) Selected independently of Alkenil.

[0326] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C8-C 20 )alkyl and (C8-C 20 ) Selected independently of Alkenil.

[0327] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C 10 -C 20 )alkyl and (C 10 -C 20 ) Selected independently of Alkenil.

[0328] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C 12 -C 20 )alkyl and (C 12 -C 20 ) Selected independently of Alkenil.

[0329] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C 14 -C 20 )alkyl and (C 14 -C 20 ) Selected independently of Alkenil.

[0330] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IV, where R 4 Each occurrence is (C 16 -C 20 )alkyl and (C 16 -C 20 ) Selected independently of Alkenil.

[0331] The present invention provides compositions comprising a compound having the structure described in formula IVa: [ka] (In the formula, R 1 is butyl, and the butyl may be substituted with one or two -OH groups. R 3 Each occurrence is independently either H or -O(CH3), R 4 Each occurrence is (C 10 -C 20 ) alkyl, (C 10 -C 20 ) Alkenyl, and (C 10 -C 20 (Selected independently of Alkinnil).

[0332] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C6-C 20 )alkyl and (C6-C 20 ) independently selected from alkenyls, and the (C6-C 20 )alkyl and (C6-C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0333] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C8-C 20 )alkyl and (C8-C 20 ) independently selected from alkenyls, and the (C8-C 20 )alkyl and (C8-C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0334] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C 10 -C 20 )alkyl and (C 10 -C 20 ) independently selected from alkenyl, and the (C 10 -C 20 )alkyl and (C 10 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0335] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C 12 -C 20 )alkyl and (C 12 -C 20 ) independently selected from alkenyl, and the (C 12 -C 20 )alkyl and (C 12 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0336] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C 14 -C 20 )alkyl and (C 14 -C 20 ) independently selected from alkenyl, and the (C 14 -C 20 )alkyl and (C 14 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0337] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C 16 -C 20 )alkyl and (C 16 -C 20 ) independently selected from alkenyl, and the (C 16 -C 20 )alkyl and (C 16 -C 20 The alkenyl group may be substituted with 1 to 6 substituents independently selected from -OH and -O(CH3).

[0338] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C6-C 20 )alkyl and (C6-C 20 ) Selected independently of Alkenil.

[0339] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C8-C 20 )alkyl and (C8-C 20 ) Selected independently of Alkenil.

[0340] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C 10 -C 20 )alkyl and (C 10 -C 20 ) Selected independently of Alkenil.

[0341] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C 12 -C 20 )alkyl and (C 12 -C 20 ) Selected independently of Alkenil.

[0342] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C 14 -C 20 )alkyl and (C 14 -C 20 ) Selected independently of Alkenil.

[0343] In some embodiments, the composition of the present invention comprises one or more Streptococcus pneumoniae antigens and a compound represented by the structure described in formula IVa, where R 4 Each occurrence is (C 16 -C 20 )alkyl and (C 16 -C 20 ) Selected independently of Alkenil.

[0344] In some embodiments, the present invention includes one or more Streptococcus pneumoniae antigens, (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, (S)-N-(5-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-yl)-5-oxopentyl)stearamide, (S)-1-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-yl)octadecane-1-one, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-yl)-5-oxopentyl)tetradecaneamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)oleamide, (9Z,12Z)-N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)octadeca-9,12-dienamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepan-1-yl)-5-oxopentyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperidine-1-yl)-5-oxopentyl)stearamide, N-(5-(3-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)azetidine-1-yl)-5-oxopentyl)stearamide, 1-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperidine-4-carboxamide, (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-octadecylcyclobutane-1-carboxamide, (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-hexadecylcyclobutane-1-carboxamide, N-(3-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-3-oxopropyl)stearamide, N-(7-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-7-oxoheptyl)stearamide, N-(3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)cyclobutyl)stearamide, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-4,4-dimethyl-5-oxopentyl)stearamide, N-(6-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-yl)-2-methyl-6-oxohexane-2-yl)stearamide, 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecyloxy)pentan-1-one, 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecylamino)pentan-1-one, N-(5-(4-(4-((5-amino-7-(butylamino)-3-methyl-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, (9Z,12Z)-N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)octadeca-9,12-dienamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)tetradecaneamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)oleamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)-4-oxobutyl)stearamide, (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)carbamate, 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperazine-1-carboxamide, 3-Stearamidopropyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-carboxylate, N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-6,6,6-trifluorohexyl)stearamide, N-(4-((4-((7-(butylamino)-5-hydroxy-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, and (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-yl)-5-oxopentyl)carbamate, The present invention provides a composition comprising a compound selected from the group consisting of, or a pharmaceutically acceptable salt thereof.

[0345] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0346] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0347] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and (S)-N-(5-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0348] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and (S)-1-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-yl)octadecane-1-one, or a pharmaceutically acceptable salt thereof.

[0349] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0350] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)tetradecanamide, or a pharmaceutically acceptable salt thereof.

[0351] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)oleamide, or a pharmaceutically acceptable salt thereof.

[0352] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and (9Z,12Z)-N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)octadeca-9,12-dienamide, or a pharmaceutically acceptable salt thereof.

[0353] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepan-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0354] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperidine-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0355] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(5-(3-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)azetidine-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0356] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and 1-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperidine-4-carboxamide, or a pharmaceutically acceptable salt thereof.

[0357] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-octadecylcyclobutan-1-carboxamide, or a pharmaceutically acceptable salt thereof.

[0358] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-hexadecylcyclobutan-1-carboxamide, or a pharmaceutically acceptable salt thereof.

[0359] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(3-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-3-oxopropyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0360] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(7-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-7-oxoheptyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0361] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)cyclobutyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0362] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-4,4-dimethyl-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0363] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(6-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-methyl-6-oxohexane-2-yl)stearamide, or a pharmaceutically acceptable salt thereof.

[0364] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecyloxy)pentan-1-one, or a pharmaceutically acceptable salt thereof.

[0365] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecylamino)pentan-1-one, or a pharmaceutically acceptable salt thereof.

[0366] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-3-methyl-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0367] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and (9Z,12Z)-N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)octadeca-9,12-dienamide, or a pharmaceutically acceptable salt thereof.

[0368] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)tetradecanamide, or a pharmaceutically acceptable salt thereof.

[0369] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)oleamide, or a pharmaceutically acceptable salt thereof.

[0370] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0371] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)-4-oxobutyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0372] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)carbamate, or a pharmaceutically acceptable salt thereof.

[0373] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperazine-1-carboxamide, or a pharmaceutically acceptable salt thereof.

[0374] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and 3-stearamidopropyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-carboxylate, or a pharmaceutically acceptable salt thereof.

[0375] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-6,6,6-trifluorohexyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0376] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and N-(4-((4-((7-(butylamino)-5-hydroxy-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide, or a pharmaceutically acceptable salt thereof.

[0377] In some embodiments, the present invention provides compositions comprising one or more Streptococcus pneumoniae antigens and (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)carbamate, or a pharmaceutically acceptable salt thereof.

[0378] Adjuvant Adjuvant-based approaches are being investigated to enhance vaccine immunogenicity and help address remaining unmet medical needs, particularly in populations highly susceptible to infection (i.e., immunocompromised, elderly, or pediatric populations).

[0379] For example, while the incidence of invasive pneumococcal disease (IPD) dramatically decreased in both children and adults after the introduction of pneumococcal conjugate vaccines, residual disease caused by persistent vaccine-type serotypes remains. Furthermore, serotype-specific immune responses vary, and the introduction of PCV with extended conjugate titers has resulted in an overall reduction in serotype-specific immune responses. Adjuvant-based strategies to enhance the immunogenicity of PCV may improve vaccine efficacy against challenging serotypes, provide a more sustained immune response, or allow for shorter dosing schedules.

[0380] Preclinical data have shown that TLR7 / 8 agonists can increase the immunogenicity of PCV. Dowling et al. reported both acceleration and enhancement of serotype-specific antibody responses, including both binding antibody titers and functional opsonin phagocytic killing, in neonatal and infant rhesus monkeys immunized with PCV13 + TLR7 / 8 agonist 3M-052 compared to PCV13 alone (PCV13 containing alum adjuvant). The addition of TLR7 / 8 agonists to PCV13 also induced Th1-polarized CRM197-specific CD4 T cells and juvenile Streptococcus pneumoniae antigen-specific B cells in infant rhesus monkeys and enhanced type II IFN and Th1-polarized cytokine production after in vitro stimulation of human neonatal umbilical cord blood (JCI Insight 2017:e91020).

[0381] Adjuvant preparations This disclosure provides an adjuvant formulation comprising 1) one or more compounds of the present invention, or (one or more) pharmaceutically acceptable salts thereof, 2) SPAN-85, 3) PS-20, and 4) squalene.

[0382] This disclosure provides an adjuvant formulation comprising: 1) a compound of the present invention or a pharmaceutically acceptable salt thereof; 2) SPAN-85; 3) PS-20 or PS-80; and 4) squalene.

[0383] This disclosure provides an adjuvant formulation comprising 1) the compound of the present invention or a pharmaceutically acceptable salt thereof, 2) SPAN-85, 3) PS-20, and 4) squalene.

[0384] General method for preparing SNE preparations Generally, SNE can be formed, for example, by first combining and mixing the components. After mixing and blending, an aqueous buffer is added and mixed with the initial compound components to form a blended emulsion mixture. In some embodiments, the blended emulsion components are first subjected to coarse homogenization, followed by fine homogenization. The resulting formulation is then subjected to a final filtration step and stored at 4°C. The solution may contain one or more compounds, one or more sorbitan-based surfactants (e.g., PS-20; PS-80; SPAN-85), and one or more terpenes (e.g., squalene) in a specific molar ratio.

[0385] Alternatively, the process for preparing the SNE of the present invention comprises four main steps: 1) solution preparation with a component mixture containing functional and non-functional components and an aqueous buffer; 2) SNE formation by stream mixing; 3) ultrafiltration; and 4) filtration.

[0386] Generally, the components are dissolved in ethanol before being sterile filtered to form a mixture. Some aqueous buffer is also prepared. The mixture and buffer streams are then combined using a T-tube or Y-mixer, then diluted immediately after the outlet and mixed with the aqueous buffer to form an SNE intermediate. The SNE intermediate is then subjected to ultrafiltration or dialysis to concentrate the material and replace the material with a suitable buffer to remove residual ethanol. After diafiltration, there is a final concentration step performed to achieve the final target concentration. The SNE bulk is then filtered through a sterile filter.

[0387] SNE preparations In some embodiments, a formulation is provided in which SNE comprises 1) about 0 to 75 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 0 to 30 mol% of SPAN-85; 3) about 0 to 30 mol% of PS-20 or PS-80; and 4) 25 to 85 mol% of squalene.

[0388] In some embodiments, a formulation is provided in which SNE comprises 1) about 0 to 50 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 0 to 10 mol% of SPAN-85; 3) about 0 to 10 mol% of PS-20 or PS-80; and 4) about 50 to 80 mol% of squalene.

[0389] In some embodiments, a formulation is provided in which SNE comprises 1) about 0 to 24 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1 to 8 mol% of SPAN-85; 3) about 1 to 8 mol% of PS-20 or PS-80; and 4) about 60 to 75 mol% of squalene.

[0390] In some embodiments, a formulation is provided in which SNE comprises 1) about 10 to 14 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1 to 4 mol% of SPAN-85; 3) about 1 to 4 mol% of PS-20 or PS-80; and 4) about 50 to 80 mol% of squalene.

[0391] In some embodiments, a formulation is provided in which SNE comprises 1) about 30 to 65 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 5 to 30 mol% of SPAN-85; 3) about 0.5 to 4 mol% of PS-20 or PS-80; and 4) about 10 to 40 mol% of squalene.

[0392] In some embodiments, the provided formulations of SNE include: 1) about 55-65 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 5-15 mol% of SPAN-85; 3) about 1-2.5 mol% of PS-20 or PS-80; and 4) about 25-35 mol% of squalene.

[0393] In some embodiments, the provided formulations of SNE include: 1) about 13 to 45 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 2 to 4 mol% of SPAN-85; 3) about 1.5 to 3 mol% of PS-20 or PS-80; and 4) about 50 to 82 mol% of squalene.

[0394] In some embodiments, a formulation is provided in which SNE comprises 1) about 13-14 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-2 mol% of SPAN-85; 3) about 1-2 mol% of PS-20 or PS-80; and 4) about 79-81 mol% of squalene.

[0395] In some embodiments, a formulation is provided in which SNE comprises: 1) about 1 to 60 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1 to 4 mol% of SPAN-85; 3) about 1 to 4 mol% of PS-20 or PS-80; and 4) about 32 to 97 mol% of squalene.

[0396] In some embodiments, the provided formulation of SNE comprises 1) about 0 to 45 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1 to 10 mol% of a nonionic surfactant; and 3) about 50 to 85 mol% of squalene. In one aspect of this embodiment, the nonionic surfactant comprises a mixture of SPAN-85 and PS-20 or PS-80.

[0397] In some embodiments, a formulation is provided in which SNE comprises 1) about 0 to 75 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 0 to 30 mol% of SPAN-85; 3) about 0 to 30 mol% of PS-20; and 4) 25 to 85 mol% of squalene.

[0398] In some embodiments, a formulation is provided in which SNE comprises 1) about 0 to 50 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 0 to 10 mol% of SPAN-85; 3) about 0 to 10 mol% of PS-20; and 4) about 50 to 80 mol% of squalene.

[0399] In some embodiments, a formulation is provided in which SNE comprises 1) about 0 to 24 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1 to 8 mol% of SPAN-85; 3) about 1 to 8 mol% of PS-20; and 4) about 60 to 75 mol% of squalene.

[0400] In some embodiments, a formulation is provided in which SNE comprises 1) about 10 to 14 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1 to 4 mol% of SPAN-85; 3) about 1 to 4 mol% of PS-20; and 4) about 50 to 80 mol% of squalene.

[0401] In some embodiments, a formulation is provided in which SNE comprises 1) about 30 to 65 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 5 to 30 mol% of SPAN-85; 3) about 0.5 to 4 mol% of PS-20; and 4) about 10 to 40 mol% of squalene.

[0402] In some embodiments, the provided formulations of SNE include: 1) about 55-65 mol% of compounds of formulas I, Ia, II, IIa, III, IIIa, IV, and IVa, or pharmaceutically acceptable salts thereof; 2) about 5-15 mol% of SPAN-85; 3) about 1-2.5 mol% of PS-20; and 4) about 25-35 mol% of squalene.

[0403] In some embodiments, the provided formulations of SNE include 1) about 13 to 45 mol% of compounds of formulas I, Ia, II, IIa, III, IIIa, IV, and IVa, or pharmaceutically acceptable salts thereof; 2) about 2 to 4 mol% of SPAN-85; 3) about 1.5 to 3 mol% of PS-20; and 4) about 50 to 82 mol% of squalene.

[0404] In some embodiments, a formulation is provided in which SNE comprises 1) about 13-14 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1-2 mol% of SPAN-85; 3) about 1-2 mol% of PS-20; and 4) about 79-81 mol% of squalene.

[0405] In some embodiments, formulations are provided in which SNE comprises: 1) about 1 to 60 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1 to 4 mol% of SPAN-85; 3) about 1 to 4 mol% of PS-20; and 4) about 32 to 97 mol% of squalene.

[0406] In some embodiments, the SNE is provided as a formulation comprising 1) about 0 to 45 mol% of a compound of formula I, Ia, II, IIa, III, IIIa, IV, and IVa, or a pharmaceutically acceptable salt thereof; 2) about 1 to 10 mol% of a nonionic surfactant; and 3) about 50 to 85 mol% of squalene. In one aspect of this embodiment, the nonionic surfactant comprises a mixture of SPAN-85 and PS-20.

[0407] In some embodiments, the formulation provided above is (N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide (compound A-1), (S)-N-(5-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide (compound A-2), (S)-1-(4-(4-((5-amino-7-((1-hydroxypentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)octadecane-1-one (compound A-3), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide (compound B-1), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)tetradecanamide (compound B-2), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)oleamide (compound B-3), (9Z,12Z)-N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)octadeca-9,12-dienamide (compound B-4), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepan-1-yl)-5-oxopentyl)stearamide (compound B-5), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperidine-1-yl)-5-oxopentyl)stearamide (compound B-6), N-(5-(3-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)azetidine-1-yl)-5-oxopentyl)stearamide (compound B-7), 1-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperidine-4-carboxamide (compound B-8), (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-octadecylcyclobutane-1-carboxamide (compound B-9), (1s,3s)-3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-N-hexadecylcyclobutane-1-carboxamide (compound B-10), N-(3-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-3-oxopropyl)stearamide (compound B-11), N-(7-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-7-oxoheptyl)stearamide (compound B-12), N-(3-(2-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)cyclobutyl)stearamide (compound B-13), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-4,4-dimethyl-5-oxopentyl)stearamide (compound B-14), N-(6-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-yl)-2-methyl-6-oxohexane-2-yl)stearamide (compound B-15), 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecyloxy)pentan-1-one (compound B-16), 1-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-(octadecylamino)pentan-1-one (compound B-17), N-(5-(4-(4-((5-amino-7-(butylamino)-3-methyl-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide (compound B-18), (9Z,12Z)-N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)octadeca-9,12-dienamide (compound C-1), N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)tetradecanamide (compound C-2), N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)oleamide (compound C-3), N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide (compound C-4), N-(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)-4-oxobutyl)stearamide (compound C-5), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(4-((4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)carbamate (compound D-1), 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-N-(3-stearamidopropyl)piperazine-1-carboxamide (compound D-2), 3-Stearamidopropyl 4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-carboxylate (compound D-3), N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-6,6,6-trifluorohexyl)stearamide (compound D-4), N-(4-((4-((7-(butylamino)-5-hydroxy-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxybenzyl)(methyl)amino)butyl)stearamide (compound D-5), and (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazine-1-yl)-5-oxopentyl)carbamate (compound D-6), It comprises a specific compound selected from, or a pharmaceutically acceptable salt thereof.

[0408] In some embodiments of the present invention, formulations are provided in which the SNE further comprises one or more additional components selected from surfactants, mixtures of surfactants, phospholipids, terpenes, terpenoids, triterpenes, or combinations thereof.

[0409] In some embodiments of the present invention, the surfactants include polyoxyethylene sorbitan ester surfactants (commonly known as Tween), particularly PS-20 and PS-80; copolymers of ethylene oxide (EO), propylene oxide (PO), and / or butylene oxide (BO), such as linear EO / PO block copolymers, sold under the trade name DOWFAX®; octoxynol, of particular interest, with a variable number of repeating ethoxy(oxy-1,2-ethanediyl) groups, such as octoxynol-9 (Triton X-100, or t-octylphenoxypolyethoxyethanol); (octylphenoxy)polyethoxyethanol (IGEPALCA-630 / NP-40); nonylphenol ethoxylates, e.g., Tergitol® NP series; and polyoxyethylene fatty ethers derived from lauryl, cetyl, stearyl, and oleyl alcohols (known as Brij surfactants), e.g., triethylene glycol mono Examples include, but are not limited to, sorbitan esters such as lauryl ether (Brij30); sorbitan trioleates (Span-85, Tween-85, or [2-[(2R,3S,4R)-4-hydroxy-3-[(Z)-octadeca-9-enoyl]oxyoxolan-2-yl]-2-[(Z)-octadeca-9-enoyl]oxyethyl](Z)-octadeca-9-enoate) and sorbitan monolaurate (commonly known as SPAN).

[0410] In some embodiments of the present invention, a mixture of surfactants, such as a PS-20 / Span85 mixture, is used. A combination of a polyoxyethylene sorbitan ester, such as polyoxyethylene sorbitan monooleate (PS-80), and an octoxynol, such as t-octylphenoxypolyethoxyethanol (Triton X-100), is also preferred. Another useful combination includes laureth-9 + polyoxyethylene sorbitan ester and / or octoxynol.

[0411] In some embodiments of the present invention, the amount of surfactant or emulsifier is 0.01 to 10 mol%, particularly about 1 to 4 mol%, of polyoxyethylene sorbitan ester (such as PS-20); 0.001 to 10 mol%, particularly about 1 to 4 mol%, of octyl- or nonylphenoxypolyoxyethanol (such as Triton X-100 or other detergents in the Triton series); w / v, particularly 0.01 to 0.1% w / v; and 0.1 to 20 mol%, preferably 0.5 to 10 mol%, particularly 1 to 4 mol%, or about 10% by mass, of polyoxyethylene ether (such as Laureth-9).

[0412] In some embodiments of the present invention, the phospholipid is selected from natural phospholipids including phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylglycerol (PG), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidic acid (phosphatidic acid salt) (PA), dipalmitoylphosphatidylcholine, monoacylphosphatidylcholine (lysoPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), N-acyl-PE, phosphoinositides, and phosphosphingolipids. Phospholipid derivatives include phosphatidic acid (DMPA, DPPA, DSPA), phosphatidylcholine (DDPC, DLPC, DMPC, DPPC, DSPC, DOPC, POPC, DEPC), phosphatidylglycerol (DMPG, DPPG, DSPG, POPG), phosphatidylethanolamine (DMPE, DPPE, DSPE DOPE), and phosphatidylserine (DOPS). Fatty acids include C14:0, palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1), linoleic acid (C18:2), linolenic acid (C18:3), and arachidonic acid (C20:4), C20:0, C22:0, as well as ethisine. In certain embodiments of the present invention, the phospholipid is phosphatidylserine, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine, 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), dilauroylphosphatidylcholine (DLPC), 1,2-dieicosenoyl-sn-glycero-3-phosphocholine, or 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC).

[0413] In some embodiments of the present invention, the terpene is selected from monoterpenes such as geraniol, terpenol, limonene, myrcene, linalool, or pinene. In some embodiments, the formulation includes humulene, farnesene, or farnesol; diterpenes such as cafestol, kahweol, sembren, or taxadiene; triterpenes such as squalene or squalante; tetraterpenes such as acyclic lycopene, monocyclic gamma-carotene, or bicyclic alpha- and beta-carotene; and sesquiterpenes consisting of polyterpenes or norisoprenoids.

[0414] Pneumococcal vaccine composition Pneumococcal vaccines or compositions are well known (e.g., PNEUMOVAX® (Merck & Co., Inc., Loway, New Jersey, USA)). Pneumococcal conjugate vaccines or compositions have been previously disclosed. See International Publication No. 2011 / 100151, International Publication No. 2019 / 139692, and International Publication No. 2020 / 131763.

[0415] Exemplary bacterial capsular polysaccharides derived from Streptococcus pneumoniae include, among others, serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20 (20A and 20B), 22F, 23A, 23B, 23F, 24F, 33F, 35B, 35F, or 38.

[0416] General method for preparing polysaccharide capsules Bacterial capsular polysaccharides, particularly those used as antigens, are suitable for use in the present invention and can be readily identified by methods for identifying immunogenic and / or antigenic polysaccharides. Exemplary bacterial capsular polysaccharides derived from Streptococcus pneumoniae include, among others, serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20 (20A and 20B), 22F, 23A, 23B, 23F, 24F, 33F, 35B, 35F, or 38.

[0417] When used herein, de-O-acetylated serotype 15B (DeOAc15B) pneumococcal polysaccharide is substantially equivalent to serotype 15C pneumococcal polysaccharide and has substantially the same NMR spectrum (data not shown). When used herein, de-O-acetylated serotype 15B pneumococcal polysaccharide and serotype 15C pneumococcal polysaccharide each have an O-acetyl content in the range of 0-5%, 0-4%, 0-3%, 0-2%, 0-1%, 0-0.5%, or 0-0.1%, or have no O-acetyl content. Spencer BL, et al. reported that pneumococcal polysaccharide 15C may be slightly O-acetylated (Spencer, BL, Let al., Clin. Vac. Immuno. (2017) 24(8):1-13). Therefore, in any embodiment of the polyvalent immunogenic compositions described herein, de-O-acetylated serotype 15B (DeOAc15B) can be used instead of serotype 15C. The de-O-acetylation process is known in the art, for example, as described in Rajam et al., Clinical and Vaccine Immunology, 2007, 14(9):1223-1227.

[0418] Polysaccharides can be purified by known techniques. However, the present invention is not limited to polysaccharides purified from natural sources, and polysaccharides may be obtained by other methods, such as total or partial synthesis. Capsular polysaccharides derived from Streptococcus pneumoniae can be prepared by standard techniques known to those skilled in the art. For example, polysaccharides can be isolated from bacteria and made to a certain size by known methods (see, for example, European Patent No. 497524 and European Patent No. 497525), preferably by microfluidization achieved using a homogenizer or by chemical hydrolysis. Streptococcus pneumoniae strains corresponding to each polysaccharide serotype may be grown in a soybean-based medium. Individual polysaccharides may then be purified through standard steps including centrifugation, precipitation, and ultrafiltration. See, for example, U.S. Patent Application Publication No. 2008 / 0286838 and U.S. Patent No. 5,847,112. Polysaccharides can be sized to reduce viscosity and / or to improve filterability and subsequent lot-to-lot consistency of the conjugated product.

[0419] Purified polysaccharides can be chemically activated to introduce functional groups that can react with carrier proteins using standard techniques. Chemical activation of polysaccharides and subsequent conjugation to carrier proteins are achieved by means described in U.S. Patents Nos. 4,365,170, 4,673,574, and 4,902,506. In short, pneumococcal polysaccharides are reacted with a periodic acid-based oxidizing agent such as sodium periodate, potassium periodate, or periodic acid to result in oxidative cleavage of adjacent hydroxyl groups to produce reactive aldehyde groups. Preferred molar equivalents of periodate (e.g., sodium periodate, sodium metaperiodate, etc.) include 0.05 to 0.5 molar equivalents (molar ratio of periodate to polysaccharide repeat units) or 0.1 to 0.5 molar equivalents. The periodate reaction can vary from 30 minutes to 24 hours depending on the diol conformation (e.g., acyclic diol, cis-diol, trans-diol) which controls the accessibility of the reactive hydroxyl group to sodium periodate.

[0420] The term "periodate" includes both periodates and periodic acid, and this term also includes metaperiodate (IO). 4- ) and orthoperiodate (IO 6- This includes both ) and various salts of periodate (e.g., sodium periodate and potassium periodate). Capsular polysaccharides can be oxidized in the presence of metaperiodate or sodium periodate (NaIO4). Furthermore, capsular polysaccharides can be oxidized in the presence of orthoperiodate or periodic acid.

[0421] Purified polysaccharides can also be linked to linkers. Once activated or linked to linkers, each capsular polysaccharide can be separately conjugated to a carrier protein to form a complex carbohydrate. Polysaccharide conjugates can be prepared by known conjugation techniques.

[0422] A polysaccharide can be bonded to a linker to form a polysaccharide-linker intermediate in which the free end of the linker is an ester group. Therefore, the linker is one in which at least one end is an ester group. The other end is selected so that it can react with the polysaccharide to form a polysaccharide-linker intermediate.

[0423] Polysaccharides can be linked to a linker using primary amine groups in the polysaccharide. In this case, the linker typically has ester groups at both ends. This allows linking to occur by reacting one of the ester groups with the primary amine group in the polysaccharide via nucleophilic acyl substitution. This reaction yields a polysaccharide-linker intermediate in which the polysaccharide is linked to the linker via an amide bond. Thus, the linker is a bifunctional linker that provides a first ester group to react with the primary amine group in the polysaccharide and a second ester group to react with the primary amine group in the support molecule. A typical linker is N-hydroxysuccinimide adipic acid diester (SIDEA).

[0424] The linkage can also be achieved indirectly, i.e., by using an additional linker that is used to derivatize the polysaccharide before linking to the linker.

[0425] Polysaccharides can be linked to an additional linker using a carbonyl group at the reducing end of the polysaccharide. This linkage involves two steps: (a1) reacting the carbonyl group with the additional linker, and (a2) reacting the free end of the additional linker with the linker. In these embodiments, the additional linker typically has primary amine groups at both ends, thereby allowing step (a1) to be carried out by reacting one of the primary amine groups with the carbonyl group in the polysaccharide by reductive amination. A primary amine group that is reactive with the carbonyl group in the polysaccharide is used. A hydrazide group or a hydroxylamino group is preferred. The same primary amine group is typically present at both ends of the additional linker, allowing for the possibility of a polysaccharide (Ps)-Ps bond. This reaction results in a polysaccharide-additional linker intermediate in which the polysaccharide is linked to the additional linker via a CN bond.

[0426] Polysaccharides can be bonded to an additional linker using different groups in the polysaccharide, particularly carboxyl groups. This bonding involves two steps: (a1) reacting the group with the additional linker, and (a2) reacting the free end of the additional linker with the linker. In this case, the additional linker typically has primary amine groups at both ends, thereby allowing step (a1) to be carried out by reacting one of the primary amine groups with a carboxyl group in the polysaccharide via EDAC activation. Primary amine groups that are reactive with the EDAC-activated carboxyl group in the polysaccharide are used. Hydrazide groups are preferred. The same primary amine group is typically present at both ends of the additional linker. This reaction results in a polysaccharide-additional linker intermediate in which the polysaccharide is bonded to the additional linker via an amide bond.

[0427] Carrier protein In certain embodiments of the present invention, CRM197 is used as a carrier protein. CRM197 is a non-toxic variant (i.e., toxoid) of diphtheria toxin. CRM197 can be isolated from a culture of Corynebacterium diphtheria strain C7(β197) grown in a caustic amino acid and yeast extract-based medium. Furthermore, CRM197 can be prepared by recombination according to the method described in U.S. Patent No. 5,614,382. Typically, CRM 197 It is purified by a combination of ultrafiltration, ammonium sulfate precipitation, and ion exchange chromatography. In some embodiments, CRM197 is prepared in Pseudomonas fluorescens using Pfenex Expression Technology (trademark) (Pfenex Inc., San Diego, California).

[0428] Other suitable carrier proteins include additional inactivated bacterial toxins such as DT (diphtheria toxoid), TT (tetanus toxoid) or TT fragment C, pertussis toxoid, cholera toxoid (e.g., described in International Publication 2004 / 083251), Escherichia coli LT, Escherichia coli ST, and exotoxin A derived from Pseudomonas aeruginosa. Other suitable carrier proteins include bacterial outer membrane proteins, such as outer membrane complex c (OMPC), porin, transferrin-binding proteins, Streptococcus pneumoniae surface protein A (PspA; see International Publication 02 / 091998), Streptococcus pneumoniae surface adhesin protein (PsaA), C5a peptidase derived from Group A or B Streptococcus, or Haemophilus influenzae protein D, Streptococcus pneumoniae pneumomolinsin (Kuo et al., 1995, Infectious Diseases). 63;2706-13), for example, ply detoxified in some form, such as dPLY-GMBS (see International Publication No. 04 / 081515) or fusions of dPLY-formol, PhtX containing PhtA, PhtB, PhtD, PhtE, and Pht proteins, such as PhtDE fusions and PhtBE fusions (see International Publication No. 01 / 98334 and International Publication No. 03 / 54007).Other proteins, such as ovalbumin, keyhole lymphed hemocyanin (KLH), bovine serum albumin (BSA), or purified protein derivatives of tuberculin (PPD), PorB (derived from Neisseria meningitidis), PD (Haemophilus influenzae protein D; see, for example, European Patent No. 0594610), or their immunologically functional equivalents, synthetic peptides (see European Patent No. 0378881 and European Patent No. 0427347), heat shock proteins (see International Publication No. 93 / 17712 and International Publication No. 94 / 03208), pertussis proteins (see International Publication No. 98 / 58668 and European Patent No. 0471177), cytokines, lymphokines, growth factors or hormones (see International Publication No. 91 / 01146), N19 protein (Baraldoi et al., 2004, Infect Immun). Artificial proteins containing multiple human CD4+ T cell epitopes from various pathogen-derived antigens (see Falugi et al., 2001, Eur J Immunol 31:3816-3824), iron uptake proteins (see International Publication No. 01 / 72337), C. difficile toxin A or B (see International Publication No. 00 / 61761), and flagellin (see Ben-Yedidia et al., 1998, Immunol Lett 64:9) can also be used as carrier proteins.

[0429] When a polyvalent vaccine is used, the second carrier can be used for one or more of the antigens in the polyvalent vaccine. The second carrier protein is preferably a protein that is non-toxic and non-reactive and can be obtained in sufficient quantity and purity. The second carrier protein also conjugates or binds to an antigen, such as a Streptococcus pneumoniae polysaccharide, to enhance the immunogenicity of the antigen. The carrier protein should follow a standard conjugation procedure. Each capsular polysaccharide that is not conjugated to the first carrier protein can be conjugated to the same second carrier protein (e.g., each capsular polysaccharide molecule is conjugated to a single carrier protein). Capsular polysaccharides that are not conjugated to the first carrier protein can be conjugated to two or more carrier proteins (each capsular polysaccharide molecule is conjugated to a single carrier protein). In such embodiments, each capsular polysaccharide of the same serotype is typically conjugated to the same carrier protein. CRM176, CRM228, CRM45 (Uchida et al., 1973, J Biol Chem 218:3838-3844); CRM9, CRM45, CRM102, CRM103, and CRM107, as well as Genetically Engineered Toxins by Nicholls and Youle, Ed: Frankel, Maecel Dekker Other DT variants can be used as a second carrier protein, such as other mutations described in Inc, 1992; deletions or mutations of Glu-148 to Asp, Gln, or Ser, and / or mutations of Ala158 to Gly, and other mutations disclosed in U.S. Patent No. 4,709,017 or U.S. Patent No. 4,950,740; mutations of at least one residue Lys516, Lys526, Phe530, and / or Lys534, and other mutations disclosed in U.S. Patent No. 5,917,017 or U.S. Patent No. 6,455,673; or fragments disclosed in U.S. Patent No. 5,843,711.

[0430] Conjugation by reductive amination The covalent bonding of polysaccharides to carrier proteins can be carried out by reductive amination, in which the amine-reactive moiety on the polysaccharide is directly bonded to the primary amine group (mainly lysine residues) of the protein. As is well known, reductive amination proceeds via a two-step mechanism. First, the Schiff base intermediate of formula R-CH=N-R' is formed by the reaction of the aldehyde group (R-CHO) on molecule 1 to the primary amine group (R'-NH2) on molecule 2. In the second step, the Schiff base is reduced to form the amino compound of formula R-CH2-NH-R'. Many reducing agents can be used, but in most cases, highly selective reducing agents such as sodium borohydride (NaCNBH3) are used because such reagents specifically reduce only the imine functional group of the Schiff base.

[0431] Since all polysaccharides have aldehyde functional groups (terminal aldehyde functional groups) at the ends of their chains, conjugation methods involving the reductive amination of polysaccharides can be applied very commonly, and if there are no other aldehyde functional groups (intrachain aldehyde functional groups) in the repeating units, such methods make it possible to obtain a conjugate in which the polysaccharide molecule is bound to a single molecule of a carrier protein.

[0432] Typical reducing agents are cyanoborohydride salts, such as sodium cyanoborohydride. While sodium cyanoborohydride is the most commonly used imine-selective reducing agent, other cyanoborohydride salts, including potassium cyanoborohydride, can be used. Differences in starting cyanide levels and residual cyanide levels during the conjugation reaction within a sodium cyanoborohydride reagent lot can lead to inconsistent conjugation performance, affecting conjugate size and conjugate Ps vs. CRM. 197 This can result in variable product attributes such as ratios. The variability of the conjugation can be reduced by controlling and / or reducing the level of free cyanide in the final reaction product.

[0433] Residual unreacted aldehydes on the polysaccharide may be reduced by adding a strong reducing agent such as sodium borohydride. Generally, the use of a strong reducing agent is preferred. However, for some polysaccharides, it is preferable to avoid this step. For example, Streptococcus pneumoniae serotype 5 contains ketone groups that readily react with strong reducing agents. In this case, it is preferable to bypass the reduction step to protect the antigenic structure of the polysaccharide.

[0434] After conjugation, the polysaccharide-protein conjugate is purified to remove excess conjugation reagent and residual free proteins and free polysaccharides by one or more of the techniques well known to those skilled in the art, including concentration / diafiltration, ultrafiltration, precipitation / elution, column chromatography, and deep filtration. See, for example, U.S. Patent No. 6,146,902. In one embodiment, the purification step is by ultrafiltration.

[0435] Pneumococcal conjugate composition The present invention provides pneumococcal conjugate compositions comprising, essentially, or alternatively comprising, any of the polysaccharide serotype-conjugate combinations described herein and / or known in the art, together with a pharmaceutically acceptable carrier and the above-mentioned SNE. The compositions may comprise, essentially consist of, or be comprised of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 or more different polysaccharide carrier protein conjugates, each of which contains a different capsular polysaccharide conjugated to a carrier protein (the carrier protein may be one specific carrier protein or two or more different carrier proteins). In one embodiment, the capsular polysaccharide is selected from at least one or more of the serotypes of Streptococcus pneumoniae: 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20 (20A or 20B), 22F, 23A, 23B, 23F, 24F, 33F, 35B, 35F, or 38. In another embodiment, the serotype is conjugated to the carrier protein CRM197.

[0436] In some embodiments, the polysaccharide contained in the polysaccharide carrier protein conjugate is selected from polysaccharides selected from the group of serotypes consisting of 1, 2, 3, 4, 5, 6A, 6B, 6C, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20 (20A and / or 20B), 22F, 23A, 23B, 23F, 24F, 33F, 35B, 35F, or 38. In other embodiments, the group of serotypes consists of 4, 6B, 9V, 14, 18C, 19F, and 23F. In other embodiments, the group of serotypes consists of 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. In other embodiments, the serotype group consists of 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F, and 33F. In other embodiments, the serotype group consists of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, and 33F. In other embodiments, the serotype group consists of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In other embodiments, the serotype group consists of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated-15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In other embodiments, the serotype group consists of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In other embodiments, the serotype group consists of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.In other embodiments, the serotype group consists of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated-15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B. In other embodiments, the serotype group consists of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

[0437] In some embodiments, the carrier protein contained in the polysaccharide carrier protein conjugate is selected from OMPC, PhtD, pLys, DT (diphtheria toxoid), TT (tetanus toxoid), TT fragment C, pertussis toxoid, cholera toxoid, and CRM197. In other embodiments, the carrier protein is CRM197.

[0438] Dosage The compositions and formulations of the present invention can be used to protect or treat people who are susceptible to infectious diseases, such as pneumococcal infections, by administering the compositions, formulations, or vaccines systemically or via mucosal routes to patients who need them. In one embodiment, the present invention provides a method for inducing an immune response to a Streptococcus pneumoniae capsular polysaccharide conjugate, comprising administering an immunologically effective amount of the immunogenic composition of the present invention to a person. In another embodiment, the present invention provides a method for vaccinating a person against a pneumococcal infection, comprising the step of administering an immunologically effective amount of the immunogenic composition, formulation, or vaccine of the present invention to a person.

[0439] The optimal amount of a component for a particular composition, formulation, or vaccine can be determined by standard studies involving observation of an appropriate immune response in the subject. For example, in another embodiment, the dosage for human vaccination is determined by extrapolation from animal studies to human data. In another embodiment, the dosage is determined empirically.

[0440] The method of the present invention can be used to prevent and / or reduce primary clinical syndromes caused by Streptococcus pneumoniae, including both invasive infections (meningitis, pneumonia, and bacteremia) and non-invasive infections (acute otitis media, sinusitis).

[0441] Administration of the compositions, formulations, or vaccines of the present invention may include one or more methods of administration: intramuscular, intraperitoneal, intradermal, or subcutaneous injection; or oral / mucous administration to the gastrointestinal, respiratory, or urogenital tract. In one embodiment, intranasal administration is used for the treatment of pneumonia or otitis media (because it can more effectively prevent nasopharyngeal carriage of Streptococcus pneumoniae and thus weaken the infection in its early stages).

[0442] The amount of conjugate in each vaccine (or composition or formulation) dose may be selected as an amount that induces an immune-protective response without significant adverse effects. Such amounts may vary depending on the serotype of Streptococcus pneumoniae. Generally, in the case of polysaccharide conjugates, each dose contains 0.1 to 100 μg of each polysaccharide, particularly 0.1 to 10 μg, and more specifically 1 to 5 μg. For example, each dose may contain 100, 150, 200, 250, 300, 400, 500 or 750 ng, or 1, 1.5, 2, 3, 4, 5, 6, 7, 7.5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 20, 22, 25, 30, 40, 50, 60, 70, 80, 90 or 100 μg.

[0443] According to any of the methods of the present invention, in one embodiment, the subject is human. In a particular embodiment, the human patient is an infant (under 1 year of age), a toddler (approximately 12 to 24 months), or a child (approximately 2 to 5 years of age). In another embodiment, the human patient is an elderly patient (>65 years of age). The compositions of the present invention are also suitable for use in older children, adolescents, and adults (e.g., 18 to 45 years of age or 18 to 65 years of age).

[0444] In one embodiment of the method of the present invention, the composition, formulation, or vaccine of the present invention is administered as a single dose. In another embodiment, the composition, formulation, or vaccine is administered two, three, or four or more times with sufficient intervals between doses. For example, the composition, formulation, or vaccine may be administered at intervals of 1, 2, 3, 4, 5, or 6 months, or any combination thereof. The immunization schedule may follow that specified for pneumococcal vaccines. For example, the routine schedule for infants and young children against invasive diseases caused by Streptococcus pneumoniae is at 2, 4, 6, and 12–15 months of age. Therefore, in a preferred embodiment, the composition, formulation, or vaccine is administered as a four-dose series at 2, 4, 6, and 12–15 months of age.

[0445] The compositions, formulations, or vaccines of the present invention may also contain one or more proteins derived from Streptococcus pneumoniae. Examples of Streptococcus pneumoniae proteins suitable for inclusion are those specified in International Patent Application Publications 02 / 083855 and 02 / 053761.

[0446] formulation In the following embodiments of the formulations, "composition" refers to a pharmaceutical composition and / or an immunogenic composition and / or a single-dose vaccine composition. In some embodiments, a composition is provided comprising one or more compounds described herein and one or more Streptococcus pneumoniae polysaccharides or one or more Streptococcus pneumoniae polysaccharide carrier protein conjugates. In some embodiments, a composition is provided comprising one or more compounds described herein and Streptococcus pneumoniae polysaccharides or Streptococcus pneumoniae polysaccharide carrier protein conjugates containing at least 7, or at least 10, or at least 13, or at least 15, or at least 20, or at least 24, or at least 27, or at least 30 or more Streptococcus pneumoniae serotypes. In some embodiments, compositions are provided comprising a compound described herein and a Streptococcus pneumoniae polysaccharide or Streptococcus pneumoniae polysaccharide carrier protein conjugate containing 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 or more Streptococcus pneumoniae serotypes. In some embodiments, compositions are provided comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F. In some embodiments, compositions are provided comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing Streptococcus pneumoniae serotypes consisting of 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. In some embodiments, compositions are provided comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing serotypes of Streptococcus pneumoniae consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F, and 33F. In some embodiments, compositions are provided comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing serotypes of Streptococcus pneumoniae consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 19F, 22F, 23F, and 33F.In some embodiments, compositions are provided comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing serotypes of Streptococcus pneumoniae consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 10A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, compositions are provided comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing serotypes of Streptococcus pneumoniae consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 10A, 12F, 10A, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, compositions are provided comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing serotypes of Streptococcus pneumoniae consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 10A, 12F, 10A, 12F, 14, 15A, 14, 15A, 15C, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B.In some embodiments, compositions are provided comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing serotypes of Streptococcus pneumoniae consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15B, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, compositions are provided comprising a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing serotypes of Streptococcus pneumoniae consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 10A, 11A, 18C, 19A, 19F, 22F, 23B, 23F, 24F, 33F, and 35B. In some embodiments, compositions are provided that include the compounds described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing serotypes of Streptococcus pneumoniae consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.In some embodiments, compositions are provided that include a compound described herein and a Streptococcus pneumoniae polysaccharide carrier protein conjugate containing serotypes of Streptococcus pneumoniae consisting of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, 15C, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B.

[0447] In some embodiments, the above composition contains an additional 1, 2, 3, 4, or 5 Streptococcus pneumoniae polysaccharide carrier protein conjugate containing other Streptococcus pneumoniae serotypes known in the art.

[0448] In some embodiments, the above composition contains an additional 1, 2, 3, 4, or 5 Streptococcus pneumoniae polysaccharide carrier protein conjugate containing other Streptococcus pneumoniae serotypes selected from groups 7C, 35F, 21, 34, 16F, 9N, 31, 17F, 23A, and 27.

[0449] In some embodiments, the above composition contains one or two additional Streptococcus pneumoniae polysaccharide carrier protein conjugates containing other Streptococcus pneumoniae serotypes selected from groups 7C, 35F, 21, and 34.

[0450] In some embodiments, the above composition contains one or two additional Streptococcus pneumoniae polysaccharide carrier protein conjugates containing other Streptococcus pneumoniae serotypes selected from groups 16F, 9N, 31, and 17F.

[0451] In some embodiments, the above composition contains one or two additional Streptococcus pneumoniae polysaccharide carrier protein conjugates containing other Streptococcus pneumoniae serotypes selected from groups 23A and 27.

[0452] In some embodiments, the above composition comprises a polysaccharide carrier protein conjugate in which the protein is the carrier protein CRM197.

[0453] In some embodiments, a composition is provided comprising about 1 μg to about 200 mg of the compound of the present invention or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which is present in a concentration of about 0.01 μg to about 100 μg per 0.5 mL of the vaccine formulation.

[0454] In some embodiments, a composition is provided comprising about 0.02 μg to about 40 mg of the compound of the present invention or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which is present in a concentration of about 0.002 μg to about 20 μg per 0.1 mL of the vaccine formulation.

[0455] In some embodiments, a composition is provided comprising about 50 μg to about 2.1 mg of the compound of the present invention or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which is present in a concentration of about 0.002 μg to about 20 μg per 0.5 mL of the vaccine formulation.

[0456] In some embodiments, a composition is provided comprising about 50 μg to about 10 mg of the compound of the present invention or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which is present in a concentration of about 0.002 μg to about 20 μg per 0.5 mL of the vaccine formulation.

[0457] In some embodiments, a composition is provided comprising about 50 μg to about 10 mg of the compound of the present invention or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which is present in a concentration of about 0.002 μg to about 20 μg per 0.5 mL of the vaccine formulation.

[0458] In some embodiments, a composition is provided comprising about 1 μg to about 200 mg of the compound of the present invention or a pharmaceutically acceptable salt thereof, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which is present in a concentration of about 0.01 μg to about 100 μg per 0.5 mL of a vaccine formulation prepared as a co-freely dried formulation.

[0459] In some embodiments, a composition is provided comprising about 1 μg to about 200 mg of the compound of the present invention or a pharmaceutically acceptable salt thereof, 1 μg to about 1 mg of aluminum in the form of APA, and at least one Streptococcus pneumoniae polysaccharide carrier protein conjugate, each of which is present in a concentration of about 0.01 μg to about 100 μg per 0.5 mL of a vaccine formulation prepared as a co-freely dried formulation.

[0460] In some embodiments, compositions are provided comprising about 0.05 μg to about 200 mg of the compound of the present invention, or a pharmaceutically acceptable salt thereof, as highlighted in the various embodiments described above, and further comprising SPAN-85, PS-80, or PS-20, and squalene. In some embodiments, compositions are provided comprising about 0.05 μg to about 200 mg of the compound of the present invention, or a pharmaceutically acceptable salt thereof, as highlighted in the various embodiments described above, and further comprising SPAN-85, PS-20, and squalene. In another embodiment, the compound is compound A-1, or A-2, or A-3. In another embodiment, the compound is compound B-1, or B-2, or B-3, or B-4, or B-5, or B-6, or B-7, or B-8, or B-9, or B-10, or B-11, or B-12, or B-13, or B-14, or B-15, or B-16, or B-17, or B-18. In another embodiment, the compound is compound C-1, or C-2, or C-3, or C-4, or C-5. In yet another embodiment, the compound is compound D-1, or D-2, or D-3, or D-4, or D-5, or D-6.

[0461] The compositions of the present invention may be administered subcutaneously, topically, orally, on the mucous membrane, intravenously, or intramuscularly. The compositions are administered in an amount sufficient to induce a protective response. The compositions can be administered via various routes, such as orally, parenterally, subcutaneously, on the mucous membrane, or intramuscularly. The dose administered may vary depending on the patient's general condition, sex, weight, and age, as well as the route of administration.

[0462] Compositions of the present invention, as highlighted in the various embodiments described above, are sometimes referred to as immunogenic compositions.

[0463] Compositions of the present invention, as highlighted in the various embodiments described above, may be referred to as vaccines or vaccine compositions.

[0464] In each of the above embodiments, the composition further comprises a Streptococcus pneumoniae polysaccharide carrier protein conjugate.

[0465] The present invention provides a method for treating or preventing pneumococcal disease by administering the above-mentioned composition.

[0466] The present invention provides the use of the above-mentioned compositions for treating or preventing pneumococcal diseases.

[0467] All publications referenced herein are incorporated by reference for the purpose of explaining and disclosing methodologies and materials that may be used in connection with the present invention. Nothing herein should be construed as an acknowledgment that the present invention has no prior rights to such disclosure by prior art.

[0468] While preferred embodiments of the present invention have been described with reference to the accompanying drawings, it should be understood that the present invention is not limited to those exact embodiments, and various modifications and changes can be made by those skilled in the art without departing from the scope or spirit of the invention as defined in the accompanying claims.

[0469] The following examples illustrate the present invention, but are not intended to limit it.

[0470] [Example 1] General methods for preparing compounds of formulas I, Ia, II, IIa, III, IIIa, IV, and IVa The compounds can be prepared from known or readily available starting materials according to methods known to those skilled in organic synthesis. Representative methods useful for preparing the compounds are described in the following examples. Alternative synthetic routes and similar structures will be apparent to those skilled in organic synthesis.

[0471] Those skilled in the art of organic synthesis will recognize that the synthesis of polycyclic and / or heterocyclic cores contained in the compounds of the present invention may require protection of certain functional groups (i.e., derivatization for chemical compatibility with specific reaction conditions). Suitable protecting groups for various functional groups of these compounds, as well as methods for their placement and removal, are well known in the field of organic chemistry. Many outlines of these methods can be found in Greene et al., Protective Groups in Organic Synthesis, Wiley-Interscience, New York, (1999).

[0472] Those skilled in organic synthesis will also recognize that one route for the synthesis of the polycyclic heterocyclic core of the compound of the present invention may be more desirable depending on the selection of accessory substituents.

[0473] Furthermore, those skilled in the art will recognize that, in some cases, the order of reactions may differ from the order presented herein in order to avoid functional group incompatibility and to adjust the synthetic route accordingly.

[0474] The preparation of polycyclic intermediates useful for creating polycyclic and / or heterocyclic cores of the compounds of the present invention is described in the literature and in official compendiums such as "Comprehensive Heterocyclic Chemistry" editions I, II, and III, published by Elsevier and edited by ARKatritzky & R.JK Taylor. The manipulation of the required substitution patterns is also described in available chemical literature summarized in official compendiums such as "Comprehensive Organic Chemistry" published by Elsevier and edited by DH R. Barton and W. DOllis; "Comprehensive Organic Functional Group Transformations" edited by ARKatritzky & R.JK Taylor; and "Comprehensive Organic Transformation" published by Wiley-CVH and edited by R.C. Larock.

[0475] The starting materials used and the intermediates prepared using the methods described in the following examples may be isolated and purified using conventional techniques, including, but not limited to, filtration, distillation, crystallization, and chromatography, if desired. Such materials may be characterized using conventional means, including physical constants and spectral data.

[0476] Those skilled in the art will be familiar with the standard formulation techniques described in published literature and textbooks such as Zheng's "Formulation and Analytical Development for Low-Dose Oral Drug Products" (Wiley, 2009, ISBN).

[0477] Preparation of compounds and intermediates The present invention will be illustrated by the following examples. For all examples, standard post-treatment and purification methods known to those skilled in the art can be used. Unless otherwise specified, all temperatures are expressed in °C (Celsius). Unless otherwise specified, all reactions are carried out at room temperature. The synthetic methodologies described herein are intended to illustrate the chemistry applicable by the use of specific examples and do not represent the scope of this disclosure.

[0478] General method Commercially available solvents, reagents, and intermediates were used as received. Non-commercial intermediates were prepared by the method described below. 1 ¹H NMR spectra are reported as ppm downfield from Me4Si, with the number, multiplicity, and binding constants of protons indicated in Hertz in parentheses. Where LC / MS data is shown, the observed parent ion is indicated. Flash column chromatography was performed using pre-packed normal-phase silica or bulk silica.

[0479] [Example 2] Method for producing intermediates of the compound of the present invention Intermediate 1-1 Preparation of compound Int.1-1 [ka] Methyl(7-(butylamino)-1H-pyrazolo[4,3-d]pyrimidine-5-yl)carbamate [ka]

[0480] Step 1: Methyl(7-hydroxy-1H-pyrazolo[4,3-d]pyrimidine-5-yl)carbamate A mixture of methyl 4-amino-1H-pyrazole-5-carboxylate (2 g, 14.17 mmol) and 1,3-bis(methoxycarbonyl)-2-methyl-2-thiopsoideura (2.92 g, 14.17 mmol) in MeOH (20 mL) was stirred, and AcOH (4.87 mL, 85 mmol) was added at ambient temperature. The resulting mixture was stirred for 16 hours, and then NaOMe (25.5 g, 142 mmol) in MeOH was added dropwise. After the addition was complete, the reaction mixture was acidified to pH 1-2 with AcOH. The resulting solid was collected by filtration, washed with CH3CN (200 mL), and then dried under vacuum to obtain the title compound. MS m / z(M+H) + Calculated value: 210.1, measured value: 210.1. 1 H-NMR(400 MHz,DMSO-d6)δ 7.83(s,1H),3.67(s,3H).

[0481] Step 2: Methyl (7-(butylamino)-1H-pyrazolo[4,3-d]pyrimidine-5-yl)carbamate Methyl(7-hydroxy-1H-pyrazolo[4,3-d]pyrimidine-5-yl)carbamate (5 g, 23.90 mmol), butan-1-amine (4.73 ml, 47.8 mmol), and DBU (3.92 mL, 26.3 mmol) were stirred in DMSO (100 mL), to which BOP (12.69 g, 28.7 mmol) was added at ambient temperature. The resulting mixture was heated at 60 °C for 3 hours. The resulting mixture was cooled to ambient temperature, then filtered, and subsequently purified directly by reverse-phase chromatography (0-50% CH3CN / water with 0.1% formic acid modifier) ​​to obtain the title compound. MS m / z(M+H) + Calculated value: 265.1, measured value: 265.1. 1 H-NMR(400 MHz,DMSO-d6)δ 12.94(br.,1H),9.89(s,1H),8.14-7.99(m,2H),3.67(s,3H),3.56-3.51(m,2H),1.66-1.58(m,2H),1.42-1.33(m,2H),0.92(t,J=7.2 Hz,3H).

[0482] Intermediate 2-1 Preparation of compound Int.2-1 [ka] 5-Azido-N-butyl-1H-pyrazolo[4,3-d]pyrimidine-7-amine [ka]

[0483] Step 1: N-butyl-5-chloro-1H-pyrazolo[4,3-d]pyrimidine-7-amine To a mixture of 5,7-dichloro-1H-pyrazolo[4,3-d]pyrimidine (3 g, 15.87 mmol) in THF (30 mL), DIEA (3.08 g, 23.81 mmol) and butane-1-amine (1.741 g, 23.81 mmol) were added at 0°C. After the addition was complete, the mixture was warmed to ambient temperature and stirred for 2 hours. The resulting mixture was diluted with water (200 mL) and then extracted with siRNA (3 × 300 mL). The combined organic extract was washed with brine (2 × 300 mL), then dried, (Na₂SO₄), and then filtered. The filtrate was concentrated to obtain the title compound, which was used directly in the next step without purification. MS m / z(M+H) + Calculated value: 226.1, measured value: 226.2.

[0484] Step 2: 5-Azido-N-butyl-1H-pyrazolo[4,3-d]pyrimidine-7-amine A mixture of N-butyl-5-chloro-1H-pyrazolo[4,3-d]pyrimidine-7-amine (3.5 g, 15.51 mmol) at ambient temperature in AcOH (6 mL) and EtOH (24 mL) was mixed with sodium azide (1.512 g, 23.26 mmol), and the mixture was then heated to 100°C for 3 hours. The resulting mixture was cooled to ambient temperature, then diluted with water (200 mL), and extracted with ethyl acetate (3 × 300 mL). The combined organic extract was washed with brine (2 × 300 mL), dried, filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-20% ethyl acetate / petroleum ether) to obtain the title compound. MS m / z (M+H) + Calculated value: 233.1, measured value: 233.1.

[0485] Using the procedure described for intermediate 2-1, the following compounds were prepared by substituting the appropriate reagent for butane-1-amine.

[0486] [Table 2]

[0487] Intermediate 2-3 Preparation of compounds Int.2-3 [ka] 5-Azido-N-butyl-3-methyl-1H-pyrazolo[4,3-d]pyrimidine-7-amine [ka]

[0488] Step 1: 5-Azido-N-butyl-3-iodo-1H-pyrazolo[4,3-d]pyrimidine-7-amine To a solution of 5-azido-N-butyl-2H-pyrazolo[4,3-d]pyrimidine-7-amine (1 g, 4.31 mmol) in DMF (30 mL), N-iodosuccinimide (1.937 g, 8.61 mmol) was added, and the mixture was then heated to 40°C. After heating overnight, the mixture was cooled to room temperature and then diluted with 10% Na2SO3 (40 mL). The resulting solid was collected by filtration, washed with water (10 mL x 3 times), and then dried under vacuum to obtain the title compound. MS m / z(M+H) + Calculated value: 359.0, measured value: 359.0.

[0489] Step 2: 5-Azido-N-butyl-3-methyl-1H-pyrazolo[4,3-d]pyrimidine-7-amine To a solution of 5-azido-N-butyl-3-iodo-2H-pyrazolo[4,3-d]pyrimidine-7-amine (1 g, 2.79 mmol) in 1,4-dioxane (10 mL) and H2O (1 mL), Pd(dppf)Cl2 (0.204 g, 0.279 mmol), K3PO4 (1.185 g, 5.58 mmol), and 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (3.51 g, 13.96 mmol) were added under N2, and the mixture was then heated overnight at 100°C. The resulting mixture was cooled to room temperature, then diluted with siRNA (30 mL), then dried with (Na2SO4), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (30-70% siRNA / petroleum ether) to obtain the title compound. MS m / z(M+H) + Calculated value: 247.1, measured value: 247.4.

[0490] Intermediate 3-1 Preparation of compound Int.3-1 [ka] tert-butyl 4-(4-(hydroxymethyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate [ka]

[0491] Step 1: tert-butyl 4-(4-formyl-3,5-dimethoxyphenyl)piperazine-1-carboxylate To a stirred mixture of 4-bromo-2,6-dimethoxybenzaldehyde (5 g, 20.40 mmol) in toluene (20 mL), tert-butylpiperazine-1-carboxylate (5.70 g, 30.6 mmol), RuPhos (1.904 g, 4.08 mmol), Pd2(dba)3 (1.868 g, 2.040 mmol), and NaOtBu (5.88 g, 61.2 mmol) were added under an argon atmosphere, and the mixture was then heated to 100°C. After 6 hours, the mixture was cooled to ambient temperature, then diluted with water (50 mL), and then extracted with siRNA (3 × 80 mL). The combined organic extract was washed with brine (3 × 50 mL), then dried with (Na2SO4), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-100% Â / DCM) to obtain the title compound. MS m / z(M+H) + Calculated value: 351.2, measured value: 351.3.

[0492] Step 2: tert-butyl 4-(4-(hydroxymethyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate To a stirred mixture of tert-butyl 4-(4-formyl-3,5-dimethoxyphenyl)piperazine-1-carboxylate (3 g, 8.56 mmol) in EtOH (10 mL), NaBH4 (0.389 g, 10.27 mmol) was added at ambient temperature. After 15 minutes, the mixture was diluted with water (15 mL) and then extracted with Et2O (3 × 50 mL). The combined organic extract was washed with brine (3 × 50 mL), then dried, (Na2SO4), and then filtered. The filtrate was then concentrated to obtain the title compound, which was used directly in the next step without purification. MS m / z (M+H) + Calculated value: 353.2, measured value: 353.3.

[0493] Intermediate 4-1 Preparation of compound Int.4-1 [ka] tert-butyl(4-(chloromethyl)-3,5-dimethoxybenzyl)(methyl)carbamate [ka]

[0494] Step 1: tert-butyl(4-formyl-3,5-dimethoxybenzyl)(methyl)carbamate 4-Bromo-2,6-dimethoxybenzaldehyde (6 g, 24.48 mmol), N-(tert-butoxycarbonyl)-N-methylglycine (9.26 g, 49.0 mmol), Ir[dF(CF3)ppy]2(dtbbpy)PF6 (0.275 g, 0.245 mmol), and [Ni(dtbbpy)(H2O)4]Cl2 (0.575 g, 1.224 mmol) were combined in DMSO (120 mL). BTMG (10 mL, 49.0 mmol) was added to this mixture. The mixture was sonicated until all solids were dissolved. The resulting mixture was uniformly divided into four screw-cap vials equipped with stirring bars. N2 was blown into each mixture for 5 minutes. The vial was capped, and the mixture was irradiated overnight using a PennOC Photoreactor® (wavelength: 420 nm; LED intensity: 100%; fan speed: 5000 rpm; stirring: 1200 rpm). The separated mixtures were combined, diluted with H2O, and then extracted with siRNA (3×). The combined organic extracts were washed with H2O and brine, dried with (MgSO4), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-50% 3:1 siRNA:EtOH / heptane) to obtain the title compound. 1 H-NMR(400 MHz,CDCl3)δ 10.47(s,1H),6.44(bs,2H),4.41(s,2H),3.88(s,6H),2.87(bs,3H),1.49(bs,9H).

[0495] Step 2: tert-butyl(4-(hydroxymethyl)-3,5-dimethoxybenzyl)(methyl)carbamate A solution of tert-butyl(4-formyl-3,5-dimethoxybenzyl)(methyl)carbamate (6.98 g, 22.56 mmol) in MeOH (100 mL) was cooled to 0°C. NaBH4 (1.03 g, 27.2 mmol) was added in fractions. After 1 hour, gas generation stopped. The cooling bath was removed and the mixture was warmed to ambient temperature. After 90 minutes, the mixture was concentrated. The residue was absorbed into DCM, filtered through a Celite® pad while washing with DCM, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-15% 3:1 siRNA:EtOH / heptane) to obtain the title compound. 1 H-NMR(400 MHz,CDCl3)δ 6.45(bs,2H),4.76(d,J=6.6 Hz,2H),4.39(s,2H),3.83(s,6H),2.86(bs,3H),1.49(s,9H).

[0496] Step 3: tert-butyl(4-(chloromethyl)-3,5-dimethoxybenzyl)(methyl)carbamate A solution of N-chlorosuccinimide (0.472 g, 3.53 mmol) in DCM (1 mL) was cooled to 0°C. Dimethyl sulfide (0.239 g, 3.85 mmol) was slowly added to this solution to obtain a precipitate. The reaction mixture was cooled to -20°C, and then a solution of tert-butyl (4-(hydroxymethyl)-3,5-dimethoxybenzyl)(methyl)carbamate (1 g, 3.21 mmol) in DCM was added dropwise. The mixture was stirred for 2 hours to reach 0°C, during which time all solid precipitate dissolved, yielding a clear solution. The solution was poured into cold salt water and then extracted with Et2O(2×). The combined organic extract was washed with cold brine, then dried with (Na2SO4), then filtered, and the filtrate was concentrated to obtain the title compound, which was used directly in the next step without purification. MS m / z(M+H) + Calculated value: 330.1, measured value: 329.2.

[0497] Intermediate 5-1 Preparation of compound Int.5-1 [ka] tert-butyl(4-oxobutyl)carbamate [ka] To a mixture of tert-butyl(4-hydroxybutyl)carbamate (1 g, 5.28 mmol) at ambient temperature in CH3CN (30 mL), IBX (1.231 g, 6.34 mmol) was added, and the mixture was then heated to 80°C. After 1 hour, the mixture was cooled to ambient temperature, then diluted with water (100 mL), and then extracted with siRNA (3 × 200 mL). The combined organic extract was washed with brine (2 × 200 mL), then dried with (Na2SO4), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-70% siRNA / petroleum ether) to obtain the title compound. 1 H-NMR (400 MHz, CDCl3): δ 3.76-3.73(m,2H),2.53-2.49(t,2H),2.03-1.97(m,2H),1.53-1.45(s,9H).

[0498] Intermediate 6-1 Preparation of compound Int.6-1 [ka] (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl(4-nitrophenyl) carbonate [ka] A mixture of (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-ol (300 mg, 0.567 mmol) and 4-nitrobenzoic anhydride hypochlorous acid (229 mg, 1.134 mmol) in DCM (4.5 mL) and pyridine (1.5 mL) was stirred at ambient temperature for 5 hours. The resulting mixture was concentrated and then purified by preparative TLC (petroleum ether:DCM = 1:1) to obtain the title compound.

[0499] Using the procedure described for intermediate 6-1, the following compounds were prepared by substituting appropriate reagents for (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-ol.

[0500] [ka]

[0501] [Table 3]

[0502] Intermediate 7-1 Preparation of compound Int.7-1 [ka] 4-Stearamidobutanoic acid [ka] A mixture of 4-aminobutanoic acid (1 g, 9.70 mmol) and Na2CO3 (1.542 g, 14.55 mmol) in THF (30 mL) and water (30 mL) was cooled to 0°C. Stearoyl chloride (2.94 g, 9.70 mmol) was added. After the addition was complete, the mixture was stirred at 25°C for 16 hours. The resulting mixture was quenched with saturated KHSO4 and then extracted with CHCl3 (500 mL). The combined organic layers were washed with brine (3 × 100 mL), dried, filtered, and the filtrate was concentrated to obtain the title compound. MS m / z (MH) - Calculated value: 368.6, measured value: 368.3.

[0503] Using the procedure described for intermediate 7-1, the following compounds were prepared by substituting appropriate reagents for 4-aminobutanoic acid.

[0504] [ka]

[0505] [Table 4]

[0506] Intermediate 8-1 Preparation of compound Int.8-1 [ka] N-(3-aminopropyl)stearamide [ka] A mixture of propane-1,3-diamine (1 g, 13.49 mmol) and pyridine (1.091 mL, 13.49 mmol) in DMF (15 mL) was cooled to 0°C. Stearoyl chloride (4.09 g, 13.49 mmol) was added. After the addition was complete, the mixture was stirred at 25°C for 16 hours. The resulting mixture was diluted with ice water to obtain a solid. The solid was collected by filtration, washed with water (3 × 100 mL), and then dried to obtain the title compound. MS m / z(M+H) + Calculated value: 341.6, measured value: 341.3.

[0507] Using the procedure described in intermediate 8-1, the following compounds were prepared by substituting appropriate reagents for propane-1,3-diamine.

[0508] [ka]

[0509] [Table 5]

[0510] Intermediate 9-1 Preparation of compound Int.9-1 [ka] 2-((1s,3s)-3-(octadecylcarbamoyl)cyclobutyl)acetic acid [ka]

[0511] Step 1: tert-butyl 2-((1s,3s)-3-(octadecylcarbamoyl)cyclobutyl) acetate (1s,3s)-3-(2-(tert-butoxy)-2-oxoethyl)cyclobutane-1-carboxylic acid (0.5 g, 2.334 mmol), HATU (1.78 g, 4.67 mmol), and DIEA (1.22 mL, 7.00 mmol) were stirred in DMF (10 mL) to which a solution of octadecane-1-amine (0.818 g, 3.03 mmol) in DMF (10 mL) was added at 25°C. After 12 hours, the mixture was diluted with water (10 mL) and then extracted with SiO2 (3 × 10 mL). The combined organic layers were washed with brine (3 × 10 mL), then dried with (Na₂SO₄), then filtered, and the filtrate was concentrated to obtain the title compound as an 80:20 mixture with tert-butyl 2-((1s,3s)-3-(hexadecylcarbamoyl)cyclobutyl) acetate. MS m / z(M+H) + Calculated value: 466.4, measured value: 466.2.

[0512] Step 2: 2-((1s,3s)-3-(octadecylcarbamoyl)cyclobutyl)acetic acid To a stirred mixture of tert-butyl 2-(3-(octadecylcarbamoyl)cyclobutyl) acetate (0.932 g, 2 mmol) in 1,4-dioxane (16 mL), 4 M HCl (4 mL) in 1,4-dioxane was added at 25 °C. After 6 hours, the mixture was diluted with ELISA and then washed with brine (3 × 30 mL). The organic layer was dried with (Na₂SO₄), then filtered, and the filtrate was concentrated to obtain the title compound as an 80:20 mixture with 2-((1s,3s)-3-(hexadecylcarbamoyl)cyclobutyl)acetic acid. MS m / z(M+H) + Calculated value: 410.4, measured value: 410.4.

[0513] Intermediate 10-1 Preparation of compound Int.10-1 [ka] 2,2-dimethyl-5-stearamidopentanoic acid [ka] A mixture of N-(5-hydroxy-4,4-dimethylpentyl)stearamide (Int. 8-3, 500 mg, 1.257 mmol) in acetone (10 mL) was mixed with chromium trioxide solution in sulfuric acid (0.691 mL, 1.383 mmol) at 0°C. After 6 hours, the mixture was diluted with water. The resulting solid was collected by filtration, washed with water, and then air-dried. MS m / z(M+H) + Calculated value: 412.4, measured value: 412.1.

[0514] Intermediate 11-1 Preparation of compound Int.11-1 [ka] 5-Methyl-5-stearamidohexanoic acid [ka]

[0515] Step 1: tert-butylmethyl glutarate A solution of 5-(tert-butoxy)-5-oxopentanoic acid (5 g, 26.6 mmol) in DCM (40 mL) and MeOH (40 mL) was cooled to 0°C. (Diazomethyl)trimethylsilane (39.8 mL, 80 mmol, 2 M in hexane) was added to this solution. After the addition was complete, the mixture was warmed to room temperature. After 3 hours, the mixture was concentrated. The crude product was subjected to silica gel chromatography (0-5% siRNA / petroleum ether) to obtain the title compound. 1H NMR(400 MHz,CDCl3)δ 3.68(s,3H),2.37(t,J=7.2 Hz,2H),2.28(t,J=7.6 Hz,2H),1.91(quin,J=7.2 Hz,2H),1.44(s,9H).

[0516] Step 2: tert-butyl 5-hydroxy-5-methylhexanoate To a solution of tert-butylmethyl glutarate (1 g, 4.94 mmol) in THF (16 mL), MeMgBr (4.12 mL, 12.36 mmol, 3 M in THF) was added at 0°C. After 2 hours, the mixture was quenched with saturated NH4Cl (25 mL), then warmed to room temperature, and extracted with siRNA (10 mL x 3). The combined organic layer was washed with brine (30 mL), then dried with (Na2SO4), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-20% siRNA / petroleum ether) to obtain the title compound. 1 H NMR(400 MHz,CDCl3)δ 2.25(t,J=7.2 Hz,2H),1.69-1.64(m,2H),1.50-1.48(m,2H),1.45(s,9H),1.23(s,6H).

[0517] Step 3: 5-methyl-5-stearamidohexanoic acid To a solution of tert-butyl 5-hydroxy-5-methylhexanoate (300 mg, 1.483 mmol) in AcOH (2 mL), stearonitrile (787 mg, 2.97 mmol) was added at room temperature, and the mixture was then cooled to 0°C. H2SO4 (0.5 mL) was added. After the addition was complete, the cooling bath was removed, and the mixture was warmed to room temperature. After stirring overnight, the mixture was quenched to pH=5 with saturated NaHCO3. The resulting mixture was extracted with RINKAN (2 mL × 3). The combined organic layers were dried over (Na2SO4), then filtered, and the filtrate was concentrated. The residue was absorbed in MeCN (5 mL). The resulting precipitate solid was collected and then recrystallized from RINKAN to obtain the title compound. MS m / z(M+H) + Calculated value: 412.7, measured value: 412.3.

[0518] Intermediate 12-1 Preparation of compound Int.12-1 [ka] 5-((tert-butoxycarbonyl)(octadecyl)amino)pentanoic acid [ka]

[0519] Step 1: Methyl 5-((tert-butoxycarbonyl)amino)pentanoate 5-((tert-butoxycarbonyl)amino)pentanoic acid (2 g, 9.21 mmol) was dissolved in MeOH (15 mL) and DCM (15 mL) and 2M (trimethylsilyl) diazomethane (23.01 mL, 46.0 mmol) in hexane at room temperature. After 1 hour, the mixture was concentrated to obtain the title compound, which was used without purification. 1 H NMR(400 MHz,CDCl3)δ 4.56(s,1H),3.68(s,3H),3.13(q,J=6.4 Hz,2H),2.34(t,J=7.2 Hz,2H),1.70-1.62(m,2H),1.49-1.56(m,2H),1.45(s,9H).

[0520] Step 2: Methyl 5-((tert-butoxycarbonyl)(octadecyl)amino)pentanoate To a solution of methyl 5-((tert-butoxycarbonyl)amino)pentanoate (1 g, 4.32 mmol) in DMF (20 mL), NaH (0.259 g, 6.49 mmol, 60% in mineral oil) was added at 0°C. After the addition was complete, the mixture was warmed to room temperature. After 30 minutes, 1-bromooctadecane (4.32 g, 12.97 mmol) was added, and the mixture was then heated to 50°C. After 2 hours, the mixture was cooled to room temperature, then diluted with water (100 mL), and then extracted with siRNA (3×). The combined organic layers were washed with brine (50 mL), then dried with (Na₂SO₄), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-20% siRNA / petroleum ether) to obtain the title compound. 1H NMR(400 MHz,CD3OD)δ 3.67-3.64(m,3H),3.23-3.15(m,4H),2.39-2.33(m,2H),1.57(d,J=4.4 Hz,6H),1.46(s,9H),1.29(s,30H),0.92-0.88(m,3H).

[0521] Step 3: 5-((tert-butoxycarbonyl)(octadecyl)amino)pentanoic acid A mixture of methyl 5-((tert-butoxycarbonyl)(octadecyl)amino)pentanoate (240 mg, 0.496 mmol) was stirred in THF (6 mL) and H2O (2 mL). LiOH monohydrate (62.5 mg, 1.488 mmol) was added at room temperature, and the mixture was then heated to 40°C. After stirring overnight, the mixture was cooled to room temperature, and the pH was adjusted to 7 with 1N HCl. The resulting mixture was extracted with siRNA (3 × 5 mL). The combined organic layers were washed with brine (3 × 5 mL), dried, filtered, and the filtrate was concentrated to obtain the title compound, which was used in the next step without purification. 1 H NMR(400 MHz,CD3OD)δ 3.23-3.14(m,4H),2.35-2.28(m,2H),1.65-1.52(m,6H),1.46(s,9H),1.29(s,30H),0.92-0.88 ppm(m,3H).

[0522] Intermediate 13-1 Preparation of compound Int.13-1 [ka] 5-(octadecyloxy)pentanoic acid [ka]

[0523] Step 1: 1-(hexa-5-en-1-yloxy)octadecane To a solution of octadecane-1-ol (5 g, 18.48 mmol) in DMF (100 mL), NaH (3.70 g, 92 mmol) (60% of mineral oil) was added in portions at room temperature. After stirring for 30 minutes, 6-bromo-1-hexene (24.70 mL, 185 mmol) was added, and the mixture was then heated to 80°C. After heating overnight, the mixture was cooled to room temperature, then diluted with water (200 mL), and then extracted with siRNA (100 mL x 3). The combined organic layers were dried (Na₂SO₄), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-100% siRNA / petroleum ether) to obtain the title compound. 1 H NMR(400 MHz,CDCl3)δ 5.82(ddt,J=17.2,10.4,6.8,6.8 Hz,1H),5.01(dq,J=17.6,1.2 Hz,1H),4.95(d,J=10.0 Hz,1H),3.45-3.36(m,4H),2.08(d,J=7.2 Hz,2H),1.65-1.55(m,5H),1.49-1.44(m,2H),1.39-1.1.17(m,29H),0.85-0.92(m,3H).

[0524] Step 2: 5-(octadecyloxy)pentanoic acid To a solution of 1-(hexa-5-en-1-yloxy)octadecane (1 g, 2.84 mmol) in MeCN (10 mL), solutions of RuCl3 (0.059 g, 0.284 mmol) and NaIO4 (2.426 g, 11.34 mmol) in H2O (10 mL) were added at room temperature. After 2 hours, the mixture was diluted with Na2SO3 (25 mL) and then extracted with siRNA (25 mL x 3). The combined organic layer was washed with 1N HCl (20 mL), saturated NaHCO3 (20 mL), and brine (20 mL), then dried to (Na2SO4), then filtered, and the filtrate was concentrated to obtain the title compound, which was used in the next step without purification. 1H NMR(400 MHz,CDCl3)δ ppm 3.39-3.44(m,4 H),2.40(t,J=7.27 Hz,2 H),1.70-1.75(m,2 H),1.65(br d,J=7.51 Hz,2 H),1.55-1.58(m,2 H),1.26(s,30 H),0.87-0.89(m,3 H).

[0525] Intermediate 14-1 Preparation of compound Int.14-1 [ka] 2-(4-(1,4-diazepan-1-yl)-2-methoxybenzyl)-5-azido-N-butyl-2H-pyrazolo[4,3-d]pyrimidine-7-amine [ka]

[0526] Step 1: tert-butyl1-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperidine-4-carboxylate To a mixture of 5-azido-2-(4-bromo-2-methoxybenzyl)-N-butyl-2H-pyrazolo[4,3-d]pyrimidine-7-amine (derived from compound B-1, step 1, 500 mg, 1.159 mmol) and tert-butyl 1,4-diazepane-1-carboxylate (302 mg, 1.507 mmol) in toluene (10 mL), RuPhos (216 mg, 0.464 mmol), Pd2(dba)3 (212 mg, 0.232 mmol), and Cs2CO3 (1133 mg, 3.48 mmol) were added under N2, and the mixture was then heated to 110°C. After heating overnight, the mixture was cooled to room temperature, then diluted with water (100 mL), and the resulting mixture was extracted with ELISA (3 × 100 mL). The combined organic layers were washed with brine (3 × 300 mL), dried, filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-52% 3:1 siRNA / petroleum ether) to obtain the title compound. MS m / z(M+H) + Calculated value: 551.7, measured value: 551.1.

[0527] Step 2: 2-(4-(1,4-diazepan-1-yl)-2-methoxybenzyl)-5-azido-N-butyl-2H-pyrazolo[4,3-d]pyrimidine-7-amine A solution of tert-butyl 4-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-1,4-diazepane-1-carboxylate (350 mg, 0.636 mmol) in DCE (2.00 mL) was cooled to 0°C. 4M HCl in 1,4-dioxane (2 mL) was added at 0°C. After 1 hour, the mixture was warmed to room temperature and then concentrated to obtain the title compound as an HCl salt, which was used without purification. MS m / z(M+H) + Calculated value: 451.3, measured value: 451.3.

[0528] Using the procedure described for intermediate 14-1, the following compounds were prepared by substituting appropriate reagents for tert-butyl 1,4-diazepane-1-carboxylate.

[0529] [ka]

[0530] [Table 6]

[0531] Intermediate 15-1 Preparation of compound Int.15-1 [ka] 5-Azido-N-butyl-2-(2-methoxy-4-(piperidine-4-yl)benzyl)-2H-pyrazolo[4,3-d]pyrimidine-7-amine [ka]

[0532] Step 1: tert-butyl4-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-3,6-dihydropyridine-1(2H)-carboxylate To a mixture of 5-azido-2-(4-bromo-2-methoxybenzyl)-N-butyl-2H-pyrazolo[4,3-d]pyrimidine-7-amine (derived from compound B-1, step 1, 400 mg, 0.927 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (430 mg, 1.391 mmol) in 1,4-dioxane (4 mL) / H2O (0.4 mL), Cs2CO3 (604 mg, 1.855 mmol) and X-PhosPdG3 (39.3 mg, 0.046 mmol) were added under N2, and the mixture was then heated to 80°C. After 2 hours, the mixture was cooled to room temperature and then diluted with ELISA (200 mL). The resulting mixture was washed with brine (3 × 100 mL), then dried with (Na₂SO₄), filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-50% siRNA / petroleum ether) to obtain the title compound. MS m / z(M+H) + Calculated value: 534.3, measured value: 534.4.

[0533] Step 2: tert-butyl 4-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperidine-1-carboxylate To a solution of tert-butyl 4-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)-3,6-dihydropyridine-1(2H)-carboxylate (290 mg, 0.543 mmol) in MeOH (10 mL), Pd / C (150 mg, 1.410 mmol) was added under N2. The resulting mixture was placed under H2 (3 × vacuum / H2) and atmosphere and stirred at room temperature. After 16 hours, the mixture was degassed (3 × vacuum / N2), then filtered and washed with MeOH, and the filtrate was concentrated to obtain the title compound. MS m / z (M+H) + Calculated value: 536.3, measured value: 536.3.

[0534] Step 3: 5-Azido-N-butyl-2-(2-methoxy-4-(piperidine-4-yl)benzyl)-2H-pyrazolo[4,3-d]pyrimidine-7-amine To a solution of tert-butyl 4-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperidine-1-carboxylate (250 mg, 0.467 mmol) in DCM (2 mL), 4 M HCl in 1,4-dioxane (2 mL) was added at room temperature. After 1 hour, the mixture was concentrated to obtain the title compound as an HCl salt, which was used without purification. MS m / z (M+H) + Calculated value: 436.3, measured value: 436.6.

[0535] Intermediate 16-1 Preparation of compound Int.16-1 [ka] 2-(4-(azetidine-3-yl)-2-methoxybenzyl)-5-azido-N-butyl-2H-pyrazolo[4,3-d]pyrimidine-7-amine [ka]

[0536] Step 1: tert-butyl3-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)azetidine-1-carboxylate To a solution of NiI2 (36.2 mg, 0.116 mmol) in DMA (1 mL), picoline imidamide hydrochloride (36.5 mg, 0.232 mmol) was added, and the mixture was then heated to 50°C. After 30 minutes, zinc powder (152 mg, 2.319 mmol) was added, followed by the addition of solutions of 5-azido-2-(4-bromo-2-methoxybenzyl)-N-butyl-2H-pyrazolo[4,3-d]pyrimidine-7-amine (derived from compound B-1, step 1, 500 mg, 1.159 mmol) and tert-butyl 3-bromoazetidine-1-carboxylate (411 mg, 1.739 mmol) in DMA (10 mL). After stirring overnight at room temperature, the mixture was directly purified by reverse-phase chromatography (CH3CN / water containing 0.1% TFA modifier) ​​to obtain the title compound. MS m / z(M+H) + Calculated value: 508.3, measured value: 508.4.

[0537] Step 2: 2-(4-(azetidine-3-yl)-2-methoxybenzyl)-5-azido-N-butyl-2H-pyrazolo[4,3-d]pyrimidine-7-amine To a solution of tert-butyl 3-(4-((5-azido-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)azetidine-1-carboxylate (60 mg, 0.118 mmol) in DCM (0.5 mL), 4 M HCl in 1,4-dioxane (0.5 mL) was added at room temperature. After 1 hour, the mixture was concentrated to obtain the title compound as an HCl salt, which was used without purification. MS m / z(M+H) + Calculated value: 408.2, measured value: 408.3.

[0538] Intermediate 17-1 Preparation of compound Int.17-1 [ka] (S)-5-azido-N-(1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)-2-(2,6-dimethoxy-4-(piperazine-1-yl)benzyl)-2H-pyrazolo[4,3-d]pyrimidine-7-amine [ka]

[0539] Step 1: (4-bromo-2,6-dimethoxyphenyl)methanol 4-bromo-2,6-dimethoxybenzaldehyde (5 g, 20.40 mmol) was dissolved in EtOH (60 mL) and NaBH4 (0.926 g, 24.48 mmol) was added at room temperature. After 1 hour, the mixture was diluted with water (50 mL) and then extracted with siRNA (30 mL x 2). The combined organic layer was washed with brine (50 mL), then dried with (Na2SO4), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (15-50% siRNA / petroleum ether) to obtain the title compound. 1H NMR(400 MHz,CDCl3)δ 6.72(s,2H),4.72(d,J=6.8 Hz,2H),3.84(s,6H),2.32(t,J=6.8 Hz,1H).

[0540] Step 2: 5-Bromo-2-(chloromethyl)-1,3-dimethoxybenzene To a solution of (4-bromo-2,6-dimethoxyphenyl)methanol (1 g, 4.05 mmol) in DCM (10 mL), SOCl2 (0.591 mL, 8.09 mmol) was added at 0°C. After the addition was complete, the mixture was warmed to room temperature and stirred overnight. The resulting mixture was concentrated to obtain the title compound, which was used in the next step without purification. 1 H NMR(400 MHz,CDCl3)δ 6.72(s,2H),4.70(s,2H),3.87(s,6H).

[0541] Step 3: (S)-2-((5-azido-2-(4-bromo-2,6-dimethoxybenzyl)-2H-pyrazolo[4,3-d]pyrimidine-7-yl)amino)pentan-1-ol (S)-2-((5-azido-2H-pyrazolo[4,3-d]pyrimidine-7-yl)amino)pentan-1-ol (Int.2-2, 800 mg, 3.05 mmol) was dissolved in DMF (15 mL) and 5-bromo-2-(chloromethyl)-1,3-dimethoxybenzene (800 mg, 3.01 mmol) and K2CO3 (1.25 g, 9.04 mmol) were added at room temperature. After stirring overnight, the mixture was diluted with water (10 mL). The resulting solid was collected by filtration and then dried under vacuum to obtain the title compound. MS m / z(M+H) + Calculated value: 491.1, measured value: 491.2. 1 H NMR(400 MHz,CD3OD)δ 8.53(s,1H),6.93(s,2H),5.64(d,J=1.6 Hz,2H),4.50-4.63(m,1H),3.88(s,6H),3.68-3.76(m,2H),1.61-1.86(m,2H),1.38-1.54(m,2H),0.97(t,J=7.2 Hz,3H).

[0542] Step 4: (S)-5-azido-2-(4-bromo-2,6-dimethoxybenzyl)-N-(1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)-2H-pyrazolo[4,3-d]pyrimidine-7-amine (S)-2-((5-azido-2-(4-bromo-2,6-dimethoxybenzyl)-2H-pyrazolo[4,3-d]pyrimidine-7-yl)amino)pentan-1-ol (900 mg, 1.832 mmol) was dissolved in DMF (15 mL), to which imidazole (374 mg, 5.50 mmol) and TBDPSCl (0.565 mL, 2.198 mmol) were added at room temperature. After stirring overnight, the mixture was diluted with water (15 mL) and then extracted with siRNA (15 mL × 2). The combined organic layer was washed with brine (20 mL), then dried with (Na2SO4), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-10% MeOH / DCM) to obtain the title compound. MS m / z (M+H) + Calculated value: 731.2, measured value: 731.2.

[0543] Step 5: Benzyl(S)-4-(4-((5-azido-7-((1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate (S)-5-azido-2-(4-bromo-2,6-dimethoxybenzyl)-N-(1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)-2H-pyrazolo[4,3-d]pyrimidine-7-amine (500 mg, 0.685 mmol) was dissolved in toluene (10 mL) and benzylpiperazine-1-carboxylate (226 mg, 1.028 mmol), RuPhos Pd G4 (58.3 mg, 0.069 mmol), and Cs2CO3 (670 mg, 2.055 mmol) were added. The mixture was purged with N2 and then heated to 90°C. After heating overnight, the mixture was cooled to room temperature, then diluted with water (10 mL), and then extracted with DCM (15 mL x 2). The combined organic layers were dried (Na2SO4), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-5% MeOH / DCM) to obtain the title compound. MS m / z(M+H) + Calculated value: 869.4, measured value: 869.4.

[0544] Step 6: (S)-5-azido-N-(1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)-2-(2,6-dimethoxy-4-(piperazin-1-yl)benzyl)-2H-pyrazolo[4,3-d]pyrimidine-7-amine Benzyl(S)-4-(4-((5-amino-7-((1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate (370 mg, 0.439 mmol) was dissolved in TFE (8 mL) and Pd / C (47 mg) was added. The mixture was degassed (3 × vacuum / N2) and then placed under an H2 (balloon) atmosphere. After 2 hours, the mixture was filtered and the filtrate was concentrated to obtain the title compound, which was used in the next step without purification. MS m / z (M+H) + Calculated value: 735.4, measured value: 735.4.

[0545] [Example 3] Preparation of compound A-1 [ka] TIFF0007872897000104.tif170170N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazin-1-yl)-5-oxopentyl)stearamide (compound A-1)

[0546] Step 1: tert-butyl4-(4-((7-(butylamino)-5-((methoxycarbonyl)amino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate To a stirred mixture of tert-butyl 4-(4-(hydroxymethyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate (Int. 3-1,13.87 g, 39.4 mmol) in benzene (80 mL), methyl (7-(butylamino)-1H-pyrazolo[4,3-d]pyrimidine-5-yl)carbamate (Int. 1-1,8 g, 30.3 mmol) and cyanomethylenetributylphosphorane (10.96 g, 45.4 mmol) were added under an argon atmosphere, and the mixture was then heated to 80°C. After 3 hours, the mixture was cooled to ambient temperature, then diluted with water (10 mL), and then extracted with ELISA (3 × 20 mL). The combined organic extract was washed with brine (3 × 10 mL), then dried with (Na₂SO₄), then filtered, and the filtrate was concentrated. The crude product was subjected to silica gel chromatography (0-100% Â / DCM) to obtain the title compound. MS m / z(M+H) + Calculated value: 599.3, measured value: 599.3.

[0547] Step 2: tert-butyl4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3,5-dimethoxyphenyl)piperazine-1-carboxylate To a stirred mixture of tert-butyl 4-(4-((7-(butylamino)-5-((methoxycarbonyl)amino)-2H-pyrazolo[4,3-d...

Claims

1. (i) A conjugate comprising 26 Streptococcus pneumoniae capsular polysaccharide carrier protein conjugates, each comprising a polysaccharide of a specific Streptococcus pneumoniae serotype conjugated to a carrier protein, wherein the Streptococcus pneumoniae serotypes consist of 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15A, de-O-acetylated 15B, 16F, 18C, 19A, 19F, 22F, 23A, 23B, 23F, 24F, 33F, and 35B, and the carrier protein is CRM197; and (ii) A compound that is N-(5-(4-(4-((5-amino-7-(butylamino)-2H-pyrazolo[4,3-d]pyrimidine-2-yl)methyl)-3-methoxyphenyl)piperazin-1-yl)-5-oxopentyl) stearamide; or a pharmaceutically acceptable salt thereof; (iii) Sorbitan trioleate (SPAN-85); (iv) Polysorbate-20 (PS-20); and (v) Squalene Nanoemulsions containing An immunogenic composition containing [the specified substance].

2. (vi) The immunogenic composition according to claim 1, further comprising a pharmaceutically acceptable carrier.

3. A pharmaceutical composition for the treatment or prevention of pneumococcal disease in human patients, comprising the immunogenic composition according to any one of claims 1 to 2.

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